Identification of Appropriate Reference Genes for qRT-PCR Analysis of Heat-Stressed Mammary Epithelial Cells in Riverine Buffaloes (Bubalus bubalis)
Identifieur interne : 000512 ( Ncbi/Merge ); précédent : 000511; suivant : 000513Identification of Appropriate Reference Genes for qRT-PCR Analysis of Heat-Stressed Mammary Epithelial Cells in Riverine Buffaloes (Bubalus bubalis)
Auteurs : Neha Kapila [Inde] ; Amit Kishore [Inde] ; Monika Sodhi [Inde] ; Ankita Sharma [Inde] ; Pawan Kumar [Inde] ; A. K. Mohanty [Inde] ; Tanushri Jerath [Inde] ; M. Mukesh [Inde]Source :
- ISRN Biotechnology [ 2090-9403 ] ; 2013.
Abstract
Gene expression studies require appropriate normalization methods for proper evaluation of reference genes. To date, not many studies have been reported on the identification of suitable reference genes in buffaloes. The present study was undertaken to determine the panel of suitable reference genes in heat-stressed buffalo mammary epithelial cells (MECs). Briefly, MEC culture from buffalo mammary gland was exposed to 42 °C for one hour and subsequently allowed to recover at 37 °C for different time intervals (from 30 m to 48 h). Three different algorithms, geNorm, NormFinder, and BestKeeper softwares, were used to evaluate the stability of 16 potential reference genes from different functional classes. Our data identified
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DOI: 10.5402/2013/735053
PubMed: 25937980
PubMed Central: 4393032
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Mohanty</name><affiliation wicri:level="1"><nlm:aff id="I3">Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana 132001, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana 132001</wicri:regionArea><wicri:noRegion>Haryana 132001</wicri:noRegion></affiliation></author><author><name sortKey="Jerath, Tanushri" sort="Jerath, Tanushri" uniqKey="Jerath T" first="Tanushri" last="Jerath">Tanushri Jerath</name><affiliation wicri:level="1"><nlm:aff id="I1">DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001</wicri:regionArea><wicri:noRegion>Haryana 132001</wicri:noRegion></affiliation></author><author><name sortKey="Mukesh, M" sort="Mukesh, M" uniqKey="Mukesh M" first="M." last="Mukesh">M. Mukesh</name><affiliation wicri:level="1"><nlm:aff id="I1">DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001</wicri:regionArea><wicri:noRegion>Haryana 132001</wicri:noRegion></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">25937980</idno><idno type="pmc">4393032</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4393032</idno><idno type="RBID">PMC:4393032</idno><idno type="doi">10.5402/2013/735053</idno><date when="2013">2013</date><idno type="wicri:Area/Pmc/Corpus">000876</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000876</idno><idno type="wicri:Area/Pmc/Curation">000876</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000876</idno><idno type="wicri:Area/Pmc/Checkpoint">000692</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000692</idno><idno type="wicri:Area/Ncbi/Merge">000512</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Identification of Appropriate Reference Genes for qRT-PCR Analysis of Heat-Stressed Mammary Epithelial Cells in Riverine Buffaloes (<italic>Bubalus bubalis</italic>)</title><author><name sortKey="Kapila, Neha" sort="Kapila, Neha" uniqKey="Kapila N" first="Neha" last="Kapila">Neha Kapila</name><affiliation wicri:level="1"><nlm:aff id="I1">DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001</wicri:regionArea><wicri:noRegion>Haryana 132001</wicri:noRegion></affiliation><affiliation wicri:level="1"><nlm:aff id="I2">Biotechnology Division, Singhania University, Jhunjhnu, Rajasthan 333515, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>Biotechnology Division, Singhania University, Jhunjhnu, Rajasthan 333515</wicri:regionArea><wicri:noRegion>Rajasthan 333515</wicri:noRegion></affiliation></author><author><name sortKey="Kishore, Amit" sort="Kishore, Amit" uniqKey="Kishore A" first="Amit" last="Kishore">Amit Kishore</name><affiliation wicri:level="1"><nlm:aff id="I1">DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001</wicri:regionArea><wicri:noRegion>Haryana 132001</wicri:noRegion></affiliation></author><author><name sortKey="Sodhi, Monika" sort="Sodhi, Monika" uniqKey="Sodhi M" first="Monika" last="Sodhi">Monika Sodhi</name><affiliation wicri:level="1"><nlm:aff id="I1">DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001</wicri:regionArea><wicri:noRegion>Haryana 132001</wicri:noRegion></affiliation></author><author><name sortKey="Sharma, Ankita" sort="Sharma, Ankita" uniqKey="Sharma A" first="Ankita" last="Sharma">Ankita Sharma</name><affiliation wicri:level="1"><nlm:aff id="I1">DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001</wicri:regionArea><wicri:noRegion>Haryana 132001</wicri:noRegion></affiliation></author><author><name sortKey="Kumar, Pawan" sort="Kumar, Pawan" uniqKey="Kumar P" first="Pawan" last="Kumar">Pawan Kumar</name><affiliation wicri:level="1"><nlm:aff id="I2">Biotechnology Division, Singhania University, Jhunjhnu, Rajasthan 333515, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>Biotechnology Division, Singhania University, Jhunjhnu, Rajasthan 333515</wicri:regionArea><wicri:noRegion>Rajasthan 333515</wicri:noRegion></affiliation></author><author><name sortKey="Mohanty, A K" sort="Mohanty, A K" uniqKey="Mohanty A" first="A. K." last="Mohanty">A. K. Mohanty</name><affiliation wicri:level="1"><nlm:aff id="I3">Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana 132001, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana 132001</wicri:regionArea><wicri:noRegion>Haryana 132001</wicri:noRegion></affiliation></author><author><name sortKey="Jerath, Tanushri" sort="Jerath, Tanushri" uniqKey="Jerath T" first="Tanushri" last="Jerath">Tanushri Jerath</name><affiliation wicri:level="1"><nlm:aff id="I1">DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001</wicri:regionArea><wicri:noRegion>Haryana 132001</wicri:noRegion></affiliation></author><author><name sortKey="Mukesh, M" sort="Mukesh, M" uniqKey="Mukesh M" first="M." last="Mukesh">M. Mukesh</name><affiliation wicri:level="1"><nlm:aff id="I1">DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001, India</nlm:aff><country xml:lang="fr">Inde</country><wicri:regionArea>DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001</wicri:regionArea><wicri:noRegion>Haryana 132001</wicri:noRegion></affiliation></author></analytic><series><title level="j">ISRN Biotechnology</title><idno type="eISSN">2090-9403</idno><imprint><date when="2013">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Gene expression studies require appropriate normalization methods for proper evaluation of reference genes. To date, not many studies have been reported on the identification of suitable reference genes in buffaloes. The present study was undertaken to determine the panel of suitable reference genes in heat-stressed buffalo mammary epithelial cells (MECs). Briefly, MEC culture from buffalo mammary gland was exposed to 42 °C for one hour and subsequently allowed to recover at 37 °C for different time intervals (from 30 m to 48 h). Three different algorithms, geNorm, NormFinder, and BestKeeper softwares, were used to evaluate the stability of 16 potential reference genes from different functional classes. Our data identified <italic>RPL4, EEF1A1,</italic> and <italic>RPS23</italic> genes to be the most appropriate reference genes that could be utilized for normalization of qPCR data in heat-stressed buffalo MECs.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Marai, I F M" uniqKey="Marai I">I. F. M. Marai</name></author><author><name sortKey="Haeeb, A A M" uniqKey="Haeeb A">A. A. M. 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<front><journal-meta><journal-id journal-id-type="nlm-ta">ISRN Biotechnol</journal-id><journal-id journal-id-type="iso-abbrev">ISRN Biotechnol</journal-id><journal-id journal-id-type="publisher-id">ISRN.BIOTECHNOLOGY</journal-id><journal-title-group><journal-title>ISRN Biotechnology</journal-title></journal-title-group><issn pub-type="epub">2090-9403</issn><publisher><publisher-name>Hindawi Publishing Corporation</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">25937980</article-id><article-id pub-id-type="pmc">4393032</article-id><article-id pub-id-type="doi">10.5402/2013/735053</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Article</subject></subj-group></article-categories><title-group><article-title>Identification of Appropriate Reference Genes for qRT-PCR Analysis of Heat-Stressed Mammary Epithelial Cells in Riverine Buffaloes (<italic>Bubalus bubalis</italic>)</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Kapila</surname><given-names>Neha</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref><xref ref-type="aff" rid="I2"><sup>2</sup></xref></contrib><contrib contrib-type="author"><name><surname>Kishore</surname><given-names>Amit</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Sodhi</surname><given-names>Monika</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Sharma</surname><given-names>Ankita</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Kumar</surname><given-names>Pawan</given-names></name><xref ref-type="aff" rid="I2"><sup>2</sup></xref></contrib><contrib contrib-type="author"><name><surname>Mohanty</surname><given-names>A. K.</given-names></name><xref ref-type="aff" rid="I3"><sup>3</sup></xref></contrib><contrib contrib-type="author"><name><surname>Jerath</surname><given-names>Tanushri</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Mukesh</surname><given-names>M.</given-names></name><xref ref-type="aff" rid="I1"><sup>1</sup></xref><xref ref-type="corresp" rid="cor1"><sup>*</sup></xref></contrib></contrib-group><aff id="I1"><sup>1</sup>DNA Fingerprinting Unit, National Bureau of Animal Genetic Resources, Karnal, Haryana 132001, India</aff><aff id="I2"><sup>2</sup>Biotechnology Division, Singhania University, Jhunjhnu, Rajasthan 333515, India</aff><aff id="I3"><sup>3</sup>Animal Biotechnology Centre, National Dairy Research Institute, Karnal, Haryana 132001, India</aff><author-notes><corresp id="cor1">*M. Mukesh: <email>mmukesh_26@hotmail.com</email></corresp><fn fn-type="other"><p>Academic Editors: R. Greiner and J. Jia</p></fn></author-notes><pub-date pub-type="collection"><year>2013</year></pub-date><pub-date pub-type="epub"><day>28</day><month>1</month><year>2013</year></pub-date><volume>2013</volume><elocation-id>735053</elocation-id><history><date date-type="received"><day>3</day><month>11</month><year>2012</year></date><date date-type="accepted"><day>23</day><month>11</month><year>2012</year></date></history><permissions><copyright-statement>Copyright © 2013 Neha Kapila et al.</copyright-statement><copyright-year>2013</copyright-year><license xlink:href="https://creativecommons.org/licenses/by/3.0/"><license-p>This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p></license></permissions><abstract><p>Gene expression studies require appropriate normalization methods for proper evaluation of reference genes. To date, not many studies have been reported on the identification of suitable reference genes in buffaloes. The present study was undertaken to determine the panel of suitable reference genes in heat-stressed buffalo mammary epithelial cells (MECs). Briefly, MEC culture from buffalo mammary gland was exposed to 42 °C for one hour and subsequently allowed to recover at 37 °C for different time intervals (from 30 m to 48 h). Three different algorithms, geNorm, NormFinder, and BestKeeper softwares, were used to evaluate the stability of 16 potential reference genes from different functional classes. Our data identified <italic>RPL4, EEF1A1,</italic> and <italic>RPS23</italic> genes to be the most appropriate reference genes that could be utilized for normalization of qPCR data in heat-stressed buffalo MECs.</p></abstract></article-meta></front><floats-group><fig id="fig1" orientation="portrait" position="float"><label>Figure 1</label><caption><p>Overall expression pattern of 16 genes evaluated in heat-stressed MECs. The data is represented as qPCR cycle threshold (Ct) values of each gene in the box and whisker diagram. The median is shown as a dashed line across the box. The boxes represent median and 1st and 3rd quartiles ranges, while whiskers indicate maximum and minimum values.</p></caption><graphic xlink:href="ISRN.BIOTECHNOLOGY2013-735053.001"></graphic></fig><fig id="fig2" orientation="portrait" position="float"><label>Figure 2</label><caption><p>Average expression stability measures (<italic>M</italic> value) for reference genes.</p></caption><graphic xlink:href="ISRN.BIOTECHNOLOGY2013-735053.002"></graphic></fig><fig id="fig3" orientation="portrait" position="float"><label>Figure 3</label><caption><p>Determination of optimal number of reference genes for normalization by calculation of pairwise variation (<italic>V</italic>) of normalization factor ratios for different number of genes.</p></caption><graphic xlink:href="ISRN.BIOTECHNOLOGY2013-735053.003"></graphic></fig><fig id="fig4" orientation="portrait" position="float"><label>Figure 4</label><caption><p>Bar plot showing gene variability in 16 candidate reference genes by NormFinder. </p></caption><graphic xlink:href="ISRN.BIOTECHNOLOGY2013-735053.004"></graphic></fig><table-wrap id="tab1" orientation="portrait" position="float"><label>Table 1</label><caption><p>General description, cellular localization, and biological functions for set of candidate genes evaluated as reference genes in the present study.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left" rowspan="1" colspan="1">Gene symbol</th><th align="left" rowspan="1" colspan="1">Description</th><th align="left" rowspan="1" colspan="1">Cellular localization</th><th align="left" rowspan="1" colspan="1">Biological function/component</th></tr></thead><tbody><tr><td align="left" rowspan="1" colspan="1"><italic>A2M </italic></td><td align="left" rowspan="1" colspan="1">Alpha 2-microglobulin </td><td align="left" rowspan="1" colspan="1">Cytoplasm</td><td align="left" rowspan="1" colspan="1">A protease inhibitor and cytokine transporter</td></tr><tr><td align="left" colspan="4" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>ACTB </italic></td><td align="left" rowspan="1" colspan="1"><italic>β</italic>-actin </td><td align="left" rowspan="1" colspan="1">Cytoplasm</td><td align="left" rowspan="1" colspan="1">Cytoskeletal structural protein, nucleotide, and ATP binding</td></tr><tr><td align="left" colspan="4" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>B2M </italic></td><td align="left" rowspan="1" colspan="1">Beta 2-microglobulin </td><td align="left" rowspan="1" colspan="1">Golgi membrane, plasma membrane, early endosome membrane, extracellular region</td><td align="left" rowspan="1" colspan="1">Cytoskeletal protein, immune response, protein binding</td></tr><tr><td align="left" colspan="4" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>EEF1A1 </italic></td><td align="left" rowspan="1" colspan="1">Eukaryotic translation elongation factor 1 alpha 1</td><td align="left" rowspan="1" colspan="1">Cytoplasm</td><td align="left" rowspan="1" colspan="1">Translation elongation factor activity</td></tr><tr><td align="left" colspan="4" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>GAPDH </italic></td><td align="left" rowspan="1" colspan="1">Glyceraldehyde 3-phosphate dehydrogenase </td><td align="left" rowspan="1" colspan="1">Plasma membrane</td><td align="left" rowspan="1" colspan="1">Glycolytic enzyme, oxidoreductase in glycolysis and gluconeogenesis</td></tr><tr><td align="left" colspan="4" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>GTP </italic></td><td align="left" rowspan="1" colspan="1">GTP-binding protein </td><td align="left" rowspan="1" colspan="1">Cytoplasm and nucleus</td><td align="left" rowspan="1" colspan="1">Biogenesis of the 60S ribosomal subunit</td></tr><tr><td align="left" colspan="4" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>HMBS </italic></td><td align="left" rowspan="1" colspan="1">Hydroxymethylbilane synthase </td><td align="left" rowspan="1" colspan="1">Cytoplasm</td><td align="left" rowspan="1" colspan="1">Heme synthesis, porphyrin metabolism, transferase activity</td></tr><tr><td align="left" colspan="4" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>HPRT1 </italic></td><td align="left" rowspan="1" colspan="1">Hypoxanthine phosphoribosyltransferase</td><td align="left" rowspan="1" colspan="1">Cytoplasm</td><td align="left" rowspan="1" colspan="1">Purine synthesis in salvage pathway</td></tr><tr><td align="left" colspan="4" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPL22 </italic></td><td align="left" rowspan="1" colspan="1">Ribosomal protein L22</td><td align="left" rowspan="1" colspan="1">Cytoplasm</td><td align="left" rowspan="1" colspan="1">Component of the 60S subunit and encodes a ribosomal protein</td></tr><tr><td align="left" colspan="4" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPL4 </italic></td><td align="left" rowspan="1" colspan="1">Ribosomal protein L4</td><td align="left" rowspan="1" colspan="1">Cytoplasm</td><td align="left" rowspan="1" colspan="1">Component of the 60S subunit and encodes a ribosomal protein </td></tr><tr><td align="left" colspan="4" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPS15 </italic></td><td align="left" rowspan="1" colspan="1">Ribosomal protein S15 </td><td align="left" rowspan="1" colspan="1">Cytoplasm</td><td align="left" rowspan="1" colspan="1">Protein synthesis/40S subribosome</td></tr><tr><td align="left" colspan="4" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPS18 </italic></td><td align="left" rowspan="1" colspan="1">Ribosomal protein S18 </td><td align="left" rowspan="1" colspan="1">Cytoplasm</td><td align="left" rowspan="1" colspan="1">Component of the 40S ribosome</td></tr><tr><td align="left" colspan="4" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPS23 </italic></td><td align="left" rowspan="1" colspan="1">Ribosomal protein S23 </td><td align="left" rowspan="1" colspan="1">Cytoplasm</td><td align="left" rowspan="1" colspan="1">Protein synthesis/40S subribosome</td></tr><tr><td align="left" colspan="4" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPS9 </italic></td><td align="left" rowspan="1" colspan="1">Ribosomal protein S9 </td><td align="left" rowspan="1" colspan="1">Cytoplasm</td><td align="left" rowspan="1" colspan="1">Protein synthesis/40S subribosome</td></tr><tr><td align="left" colspan="4" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>UBC </italic></td><td align="left" rowspan="1" colspan="1">Ubiquitin C</td><td align="left" rowspan="1" colspan="1">Cytoplasm and nucleus</td><td align="left" rowspan="1" colspan="1">Protein degradation</td></tr><tr><td align="left" colspan="4" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>UXT </italic></td><td align="left" rowspan="1" colspan="1">Ubiquitously expressed transcript </td><td align="left" rowspan="1" colspan="1">Cytoplasm and nucleus</td><td align="left" rowspan="1" colspan="1">Transcriptional activation, ATP binding, microtubule binding, unfolded protein binding</td></tr></tbody></table></table-wrap><table-wrap id="tab2" orientation="portrait" position="float"><label>Table 2</label><caption><p>Gene name, GenBank accession numbers, primer sequences, annealing temperature (<italic>T</italic><sub><italic>a</italic></sub>), amplicon length, and PCR efficiency for the studied genes.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left" rowspan="1" colspan="1">Genes</th><th align="center" rowspan="1" colspan="1">Accession number</th><th align="center" rowspan="1" colspan="1">Primers 5′-3′ (forward, reverse)</th><th align="center" rowspan="1" colspan="1"><italic>T</italic><sub><italic>a</italic></sub></th><th align="center" rowspan="1" colspan="1">Amplicon size (bp)</th><th align="center" rowspan="1" colspan="1">PCR efficiency (%)<sup>4</sup></th></tr></thead><tbody><tr><td align="left" rowspan="1" colspan="1"><italic>A2M</italic><sup>1</sup></td><td align="center" rowspan="1" colspan="1">CR452243</td><td align="center" rowspan="1" colspan="1">CACCCAGGACACAGTGGTAGC<break></break>CCCTGAAGACTGGATGGTCAC</td><td align="center" rowspan="1" colspan="1">60°C</td><td align="center" rowspan="1" colspan="1">103</td><td align="center" rowspan="1" colspan="1">97.20</td></tr><tr><td align="left" colspan="6" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>ACTB</italic><sup>1</sup></td><td align="center" rowspan="1" colspan="1">AY141970</td><td align="center" rowspan="1" colspan="1">GCGTGGCTACAGCTTCACC<break></break>TTGATGTCACGGACGATTTC</td><td align="center" rowspan="1" colspan="1">60°C</td><td align="center" rowspan="1" colspan="1">56</td><td align="center" rowspan="1" colspan="1">90.70</td></tr><tr><td align="left" colspan="6" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>B2M </italic></td><td align="center" rowspan="1" colspan="1">NM_173893</td><td align="center" rowspan="1" colspan="1">CTGCTATGTGTATGGGTTCC<break></break>GGAGTGAACTCAGCGTG</td><td align="center" rowspan="1" colspan="1">60°C</td><td align="center" rowspan="1" colspan="1">101</td><td align="center" rowspan="1" colspan="1">131.37</td></tr><tr><td align="left" colspan="6" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>EEF1A1 </italic></td><td align="center" rowspan="1" colspan="1">BC105315</td><td align="center" rowspan="1" colspan="1">CATCCCAGGCTGACTGTGC<break></break>TGTAAGCCAAAAGGGCATGC</td><td align="center" rowspan="1" colspan="1">60°C</td><td align="center" rowspan="1" colspan="1">101</td><td align="center" rowspan="1" colspan="1">115.50</td></tr><tr><td align="left" colspan="6" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>GAPDH</italic><sup>1</sup></td><td align="center" rowspan="1" colspan="1">BC102589</td><td align="center" rowspan="1" colspan="1">TGGAAAGGCCATCACCATCT<break></break>CCCACTTGATGTTGGCAG</td><td align="center" rowspan="1" colspan="1">60°C</td><td align="center" rowspan="1" colspan="1">60</td><td align="center" rowspan="1" colspan="1">96.64</td></tr><tr><td align="left" colspan="6" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>GTP</italic><sup>1</sup></td><td align="center" rowspan="1" colspan="1">AK074976</td><td align="center" rowspan="1" colspan="1">CTTGGAATCCGAGGAGCCA<break></break>CCTGGGATCACCAGAGCTGT</td><td align="center" rowspan="1" colspan="1">60°C</td><td align="center" rowspan="1" colspan="1">101</td><td align="center" rowspan="1" colspan="1">102.91</td></tr><tr><td align="left" colspan="6" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>HMBS</italic><sup>2</sup></td><td align="center" rowspan="1" colspan="1">BC112573.1</td><td align="center" rowspan="1" colspan="1">CTTTGGAGAGGAATGAAGTGG<break></break>AATGGTGAAGCCAGGAGGAA</td><td align="center" rowspan="1" colspan="1">60°C</td><td align="center" rowspan="1" colspan="1">80</td><td align="center" rowspan="1" colspan="1">101.4</td></tr><tr><td align="left" colspan="6" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>HPRTI</italic><sup>3</sup></td><td align="center" rowspan="1" colspan="1">BC103248</td><td align="center" rowspan="1" colspan="1">GAGAAGTCCGAGTTGAGTTTGGAA<break></break>GGCTCGTAGTGCAAATGAAGAGT</td><td align="center" rowspan="1" colspan="1">64°C</td><td align="center" rowspan="1" colspan="1">190</td><td align="center" rowspan="1" colspan="1">99.70</td></tr><tr><td align="left" colspan="6" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPL22</italic><sup>1</sup></td><td align="center" rowspan="1" colspan="1">BC114880</td><td align="center" rowspan="1" colspan="1">AAGATGGCGCCGAAGAAAG <break></break>TTTCCCGAATCAAAAATTCCA</td><td align="center" rowspan="1" colspan="1">60°C</td><td align="center" rowspan="1" colspan="1">101</td><td align="center" rowspan="1" colspan="1">102.73</td></tr><tr><td align="left" colspan="6" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPL4 </italic></td><td align="center" rowspan="1" colspan="1">NM_001014894</td><td align="center" rowspan="1" colspan="1"> TTGGAAACATGTGTCGTGGG<break></break>GCAGATGGCGTATCGCTTCT</td><td align="center" rowspan="1" colspan="1">60°C</td><td align="center" rowspan="1" colspan="1">101</td><td align="center" rowspan="1" colspan="1">93.72</td></tr><tr><td align="left" colspan="6" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPS15A </italic></td><td align="center" rowspan="1" colspan="1">BC108231</td><td align="center" rowspan="1" colspan="1"> GAATGGTGCGCATGAATGTC<break></break>GACTTTGGAGCACGGCCTAA</td><td align="center" rowspan="1" colspan="1">60°C</td><td align="center" rowspan="1" colspan="1">101</td><td align="center" rowspan="1" colspan="1">119.08</td></tr><tr><td align="left" colspan="6" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RS18 </italic></td><td align="center" rowspan="1" colspan="1">DQ222453.1</td><td align="center" rowspan="1" colspan="1">TTGCCTTTGCCATCACTG<break></break>CTTGTATTGGCGTGGATT</td><td align="center" rowspan="1" colspan="1">60°C</td><td align="center" rowspan="1" colspan="1">158</td><td align="center" rowspan="1" colspan="1">129.97</td></tr><tr><td align="left" colspan="6" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPS23</italic><sup>1</sup></td><td align="center" rowspan="1" colspan="1">BC102049</td><td align="center" rowspan="1" colspan="1">CCCAATGATGGTTGCTTGAA<break></break>CGGACTCCAGGAATGTCACC</td><td align="center" rowspan="1" colspan="1">60°C</td><td align="center" rowspan="1" colspan="1">101</td><td align="center" rowspan="1" colspan="1">115.5</td></tr><tr><td align="left" colspan="6" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPS9</italic><sup>1</sup></td><td align="center" rowspan="1" colspan="1">DT860044</td><td align="center" rowspan="1" colspan="1">CCTCGACCAAGAGCTGAAG<break></break>CCTCCAGACCTCACGTTTGTTC</td><td align="center" rowspan="1" colspan="1">60°C</td><td align="center" rowspan="1" colspan="1">54</td><td align="center" rowspan="1" colspan="1">96.45</td></tr><tr><td align="left" colspan="6" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>UBC</italic><sup>2</sup></td><td align="center" rowspan="1" colspan="1">BE668033</td><td align="center" rowspan="1" colspan="1">TCCCTACCTGCATCATGTGC<break></break>GGAATTTGGGCCAGTGCTC</td><td align="center" rowspan="1" colspan="1">59°C</td><td align="center" rowspan="1" colspan="1">71</td><td align="center" rowspan="1" colspan="1">104.71</td></tr><tr><td align="left" colspan="6" rowspan="1"><hr></hr></td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>UXT</italic><sup>1</sup></td><td align="center" rowspan="1" colspan="1">CR452243</td><td align="center" rowspan="1" colspan="1">TGTGGCCCTTGGATATGGTT<break></break>GGTTGTCGCTGAGCTCTGTG</td><td align="center" rowspan="1" colspan="1">60°C</td><td align="center" rowspan="1" colspan="1">101</td><td align="center" rowspan="1" colspan="1">99.74</td></tr></tbody></table><table-wrap-foot><fn><p><sup>1</sup>Bionaz and Loor [<xref rid="B19" ref-type="bibr">19</xref>],<sup>2</sup>Pérez et al. [<xref rid="B39" ref-type="bibr">32</xref>], and <sup>3</sup>Hernandez et al. [<xref rid="B40" ref-type="bibr">33</xref>].</p></fn><fn><p><sup>4</sup>qPCR efficiencies for each primer pair were calculated from six-point standard curves using fivefold dilution series of pooled cDNA from control and heat-stressed samples.</p></fn></table-wrap-foot></table-wrap><table-wrap id="tab3" orientation="portrait" position="float"><label>Table 3</label><caption><p>Ranking of reference genes based on their expression stability.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left" rowspan="1" colspan="1">Ranking order</th><th align="center" rowspan="1" colspan="1">Genes</th><th align="center" rowspan="1" colspan="1"><italic>M</italic> value</th></tr></thead><tbody><tr><td align="left" rowspan="1" colspan="1">1</td><td align="center" rowspan="1" colspan="1"><italic>EEF1A1 </italic></td><td align="center" rowspan="1" colspan="1">0.089</td></tr><tr><td align="left" rowspan="1" colspan="1">2</td><td align="center" rowspan="1" colspan="1"><italic>RPL4 </italic></td><td align="center" rowspan="1" colspan="1">0.089</td></tr><tr><td align="left" rowspan="1" colspan="1">3</td><td align="center" rowspan="1" colspan="1"><italic>RPS23 </italic></td><td align="center" rowspan="1" colspan="1">0.109</td></tr><tr><td align="left" rowspan="1" colspan="1">4</td><td align="center" rowspan="1" colspan="1"><italic>GTP </italic></td><td align="center" rowspan="1" colspan="1">0.130</td></tr><tr><td align="left" rowspan="1" colspan="1">5</td><td align="center" rowspan="1" colspan="1"><italic>UXT </italic></td><td align="center" rowspan="1" colspan="1">0.142</td></tr><tr><td align="left" rowspan="1" colspan="1">6</td><td align="center" rowspan="1" colspan="1"><italic>RPS9 </italic></td><td align="center" rowspan="1" colspan="1">0.151</td></tr><tr><td align="left" rowspan="1" colspan="1">7</td><td align="center" rowspan="1" colspan="1"><italic>RPS15A </italic></td><td align="center" rowspan="1" colspan="1">0.157</td></tr><tr><td align="left" rowspan="1" colspan="1">8</td><td align="center" rowspan="1" colspan="1"><italic>HMBS </italic></td><td align="center" rowspan="1" colspan="1">0.164</td></tr><tr><td align="left" rowspan="1" colspan="1">9</td><td align="center" rowspan="1" colspan="1"><italic>B2M </italic></td><td align="center" rowspan="1" colspan="1">0.174</td></tr><tr><td align="left" rowspan="1" colspan="1">10</td><td align="center" rowspan="1" colspan="1"><italic>HPRT1 </italic></td><td align="center" rowspan="1" colspan="1">0.182</td></tr><tr><td align="left" rowspan="1" colspan="1">11</td><td align="center" rowspan="1" colspan="1"><italic>UBC </italic></td><td align="center" rowspan="1" colspan="1">0.201</td></tr><tr><td align="left" rowspan="1" colspan="1">12</td><td align="center" rowspan="1" colspan="1"><italic>RS18 </italic></td><td align="center" rowspan="1" colspan="1">0.219</td></tr><tr><td align="left" rowspan="1" colspan="1">13</td><td align="center" rowspan="1" colspan="1"><italic>GAPDH </italic></td><td align="center" rowspan="1" colspan="1">0.236</td></tr><tr><td align="left" rowspan="1" colspan="1">14</td><td align="center" rowspan="1" colspan="1"><italic>ACTB </italic></td><td align="center" rowspan="1" colspan="1">0.251</td></tr><tr><td align="left" rowspan="1" colspan="1">15</td><td align="center" rowspan="1" colspan="1"><italic>RPL22 </italic></td><td align="center" rowspan="1" colspan="1">0.305</td></tr><tr><td align="left" rowspan="1" colspan="1">16</td><td align="center" rowspan="1" colspan="1"><italic>A2M </italic></td><td align="center" rowspan="1" colspan="1">0.415</td></tr></tbody></table></table-wrap><table-wrap id="tab4" orientation="portrait" position="float"><label>Table 4</label><caption><p>Standard deviation (SD) and accumulated standard deviation (Acc. SD) for each gene analyzed through NormFinder.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left" rowspan="1" colspan="1">Genes</th><th align="center" rowspan="1" colspan="1">SD</th><th align="center" rowspan="1" colspan="1">Acc. SD</th></tr></thead><tbody><tr><td align="left" rowspan="1" colspan="1"><italic>RPL4 </italic></td><td align="center" rowspan="1" colspan="1">0.0115</td><td align="center" rowspan="1" colspan="1">0.0115</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>EEF1A1 </italic></td><td align="center" rowspan="1" colspan="1">0.0446</td><td align="center" rowspan="1" colspan="1">0.023</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>GTP </italic></td><td align="center" rowspan="1" colspan="1">0.1029</td><td align="center" rowspan="1" colspan="1">0.0376</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>UXT </italic></td><td align="center" rowspan="1" colspan="1">0.1056</td><td align="center" rowspan="1" colspan="1">0.0386</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPS23 </italic></td><td align="center" rowspan="1" colspan="1">0.1397</td><td align="center" rowspan="1" colspan="1">0.0416</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPS9 </italic></td><td align="center" rowspan="1" colspan="1">0.1457</td><td align="center" rowspan="1" colspan="1">0.0424</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>B2M </italic></td><td align="center" rowspan="1" colspan="1">0.1578</td><td align="center" rowspan="1" colspan="1">0.0427</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPS15A </italic></td><td align="center" rowspan="1" colspan="1">0.176 </td><td align="center" rowspan="1" colspan="1">0.0434</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>UBC </italic></td><td align="center" rowspan="1" colspan="1">0.1762</td><td align="center" rowspan="1" colspan="1">0.0432</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>HMBS </italic></td><td align="center" rowspan="1" colspan="1">0.2081</td><td align="center" rowspan="1" colspan="1">0.0441</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>HPRT1 </italic></td><td align="center" rowspan="1" colspan="1">0.2147</td><td align="center" rowspan="1" colspan="1">0.0446</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>GAPDH </italic></td><td align="center" rowspan="1" colspan="1">0.2591</td><td align="center" rowspan="1" colspan="1">0.0463</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>ACTB </italic></td><td align="center" rowspan="1" colspan="1">0.2735</td><td align="center" rowspan="1" colspan="1">0.0476</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RS18 </italic></td><td align="center" rowspan="1" colspan="1">0.3278</td><td align="center" rowspan="1" colspan="1">0.05</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPL22 </italic></td><td align="center" rowspan="1" colspan="1">0.6443</td><td align="center" rowspan="1" colspan="1">0.0634</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>A2M </italic></td><td align="center" rowspan="1" colspan="1">1.1554</td><td align="center" rowspan="1" colspan="1">0.0936</td></tr></tbody></table></table-wrap><table-wrap id="tab5" orientation="portrait" position="float"><label>Table 5</label><caption><p>Parameters based cycle point (CP) values for the 10 most stable housekeeping genes.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left" rowspan="1" colspan="1"></th><th align="center" rowspan="1" colspan="1"><italic>EEF1A1 </italic></th><th align="center" rowspan="1" colspan="1"><italic>RPL4 </italic></th><th align="center" rowspan="1" colspan="1"><italic>RPS23 </italic></th><th align="center" rowspan="1" colspan="1"><italic>GTP </italic></th><th align="center" rowspan="1" colspan="1"><italic>UXT </italic></th><th align="center" rowspan="1" colspan="1"><italic>RPS9 </italic></th><th align="center" rowspan="1" colspan="1"><italic>RPS15A </italic></th><th align="center" rowspan="1" colspan="1"><italic>HMBS </italic></th><th align="center" rowspan="1" colspan="1"><italic>B2M </italic></th><th align="center" rowspan="1" colspan="1"><italic>HPRT1 </italic></th></tr></thead><tbody><tr><td align="left" rowspan="1" colspan="1"><italic>n</italic></td><td align="center" rowspan="1" colspan="1">9</td><td align="center" rowspan="1" colspan="1">9</td><td align="center" rowspan="1" colspan="1">9</td><td align="center" rowspan="1" colspan="1">9</td><td align="center" rowspan="1" colspan="1">9</td><td align="center" rowspan="1" colspan="1">9</td><td align="center" rowspan="1" colspan="1">9</td><td align="center" rowspan="1" colspan="1">9</td><td align="center" rowspan="1" colspan="1">9</td><td align="center" rowspan="1" colspan="1">9</td></tr><tr><td align="left" rowspan="1" colspan="1">GM [CP]</td><td align="center" rowspan="1" colspan="1">23.64</td><td align="center" rowspan="1" colspan="1">22.02</td><td align="center" rowspan="1" colspan="1">19.81</td><td align="center" rowspan="1" colspan="1">24.94</td><td align="center" rowspan="1" colspan="1">25.90</td><td align="center" rowspan="1" colspan="1">19.15</td><td align="center" rowspan="1" colspan="1">19.15</td><td align="center" rowspan="1" colspan="1">25.81</td><td align="center" rowspan="1" colspan="1">22.07</td><td align="center" rowspan="1" colspan="1">22.57</td></tr><tr><td align="left" rowspan="1" colspan="1">AM [CP]</td><td align="center" rowspan="1" colspan="1">23.64</td><td align="center" rowspan="1" colspan="1">22.02</td><td align="center" rowspan="1" colspan="1">19.81</td><td align="center" rowspan="1" colspan="1">24.94</td><td align="center" rowspan="1" colspan="1">25.90</td><td align="center" rowspan="1" colspan="1">19.54</td><td align="center" rowspan="1" colspan="1">19.15</td><td align="center" rowspan="1" colspan="1">25.81</td><td align="center" rowspan="1" colspan="1">22.07</td><td align="center" rowspan="1" colspan="1">22.57</td></tr><tr><td align="left" rowspan="1" colspan="1">min [CP]</td><td align="center" rowspan="1" colspan="1">23.31</td><td align="center" rowspan="1" colspan="1">21.79</td><td align="center" rowspan="1" colspan="1">19.42</td><td align="center" rowspan="1" colspan="1">24.54</td><td align="center" rowspan="1" colspan="1">25.70</td><td align="center" rowspan="1" colspan="1">10.27</td><td align="center" rowspan="1" colspan="1">18.80</td><td align="center" rowspan="1" colspan="1">25.31</td><td align="center" rowspan="1" colspan="1">21.86</td><td align="center" rowspan="1" colspan="1">22.14</td></tr><tr><td align="left" rowspan="1" colspan="1">max [CP]</td><td align="center" rowspan="1" colspan="1">23.91</td><td align="center" rowspan="1" colspan="1">22.30</td><td align="center" rowspan="1" colspan="1">20.02</td><td align="center" rowspan="1" colspan="1">25.27</td><td align="center" rowspan="1" colspan="1">26.30</td><td align="center" rowspan="1" colspan="1">20.87</td><td align="center" rowspan="1" colspan="1">19.43</td><td align="center" rowspan="1" colspan="1">26.06</td><td align="center" rowspan="1" colspan="1">22.37</td><td align="center" rowspan="1" colspan="1">23.00</td></tr><tr><td align="left" rowspan="1" colspan="1">SD [±CP]</td><td align="center" rowspan="1" colspan="1">0.13</td><td align="center" rowspan="1" colspan="1">0.11</td><td align="center" rowspan="1" colspan="1">0.15</td><td align="center" rowspan="1" colspan="1">0.16</td><td align="center" rowspan="1" colspan="1">0.12</td><td align="center" rowspan="1" colspan="1">2.06</td><td align="center" rowspan="1" colspan="1">0.13</td><td align="center" rowspan="1" colspan="1">0.18</td><td align="center" rowspan="1" colspan="1">0.11</td><td align="center" rowspan="1" colspan="1">0.20</td></tr><tr><td align="left" rowspan="1" colspan="1">CV [%CP]</td><td align="center" rowspan="1" colspan="1">0.57</td><td align="center" rowspan="1" colspan="1">0.52</td><td align="center" rowspan="1" colspan="1">0.76</td><td align="center" rowspan="1" colspan="1">0.64</td><td align="center" rowspan="1" colspan="1">0.47</td><td align="center" rowspan="1" colspan="1">10.54</td><td align="center" rowspan="1" colspan="1">0.67</td><td align="center" rowspan="1" colspan="1">0.71</td><td align="center" rowspan="1" colspan="1">0.49</td><td align="center" rowspan="1" colspan="1">0.89</td></tr><tr><td align="left" rowspan="1" colspan="1">min [x-fold]</td><td align="center" rowspan="1" colspan="1">−1.26</td><td align="center" rowspan="1" colspan="1">−1.18</td><td align="center" rowspan="1" colspan="1">−1.31</td><td align="center" rowspan="1" colspan="1">−1.32</td><td align="center" rowspan="1" colspan="1">−1.15</td><td align="center" rowspan="1" colspan="1">−470.28</td><td align="center" rowspan="1" colspan="1">−1.28</td><td align="center" rowspan="1" colspan="1">−1.42</td><td align="center" rowspan="1" colspan="1">−1.16</td><td align="center" rowspan="1" colspan="1">−1.34</td></tr><tr><td align="left" rowspan="1" colspan="1">max [x-fold]</td><td align="center" rowspan="1" colspan="1">1.21</td><td align="center" rowspan="1" colspan="1">1.21</td><td align="center" rowspan="1" colspan="1">1.16</td><td align="center" rowspan="1" colspan="1">1.25</td><td align="center" rowspan="1" colspan="1">1.31</td><td align="center" rowspan="1" colspan="1">3.30</td><td align="center" rowspan="1" colspan="1">1.21</td><td align="center" rowspan="1" colspan="1">1.19</td><td align="center" rowspan="1" colspan="1">1.23</td><td align="center" rowspan="1" colspan="1">1.35</td></tr><tr><td align="left" rowspan="1" colspan="1">SD [±x-fold]</td><td align="center" rowspan="1" colspan="1">1.10</td><td align="center" rowspan="1" colspan="1">1.08</td><td align="center" rowspan="1" colspan="1">1.11</td><td align="center" rowspan="1" colspan="1">1.12</td><td align="center" rowspan="1" colspan="1">1.09</td><td align="center" rowspan="1" colspan="1">4.17</td><td align="center" rowspan="1" colspan="1">1.09</td><td align="center" rowspan="1" colspan="1">1.13</td><td align="center" rowspan="1" colspan="1">1.08</td><td align="center" rowspan="1" colspan="1">1.15</td></tr></tbody></table><table-wrap-foot><fn><p><italic>n</italic>: number of samples; GM [CP]: geometric mean of cycling point; AM [CP]: arithmetic mean of CP; min [CP] and max [CP]: extreme values of CP; SD [±CP]: standard deviation of the CP; CV [%CP]: coefficient of variation expressed as a percentage on the CP values; min [x-fold] and max [x-fold]: extreme values of expression levels expressed as absolute x-fold over or under coefficient; SD [±x-fold]: standard deviation of the absolute regulation coefficients.</p></fn></table-wrap-foot></table-wrap><table-wrap id="tab6" orientation="portrait" position="float"><label>Table 6</label><caption><p>Repeated pairwise correlation analysis among genes and with BestKeeper index (BI).</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="left" rowspan="1" colspan="1">Genes</th><th align="center" rowspan="1" colspan="1">versus</th><th align="center" rowspan="1" colspan="1"><italic>EEF1A1 </italic></th><th align="center" rowspan="1" colspan="1"><italic>RPL4 </italic></th><th align="center" rowspan="1" colspan="1"><italic>RPS23 </italic></th><th align="center" rowspan="1" colspan="1"><italic>GTP </italic></th><th align="center" rowspan="1" colspan="1"><italic>UXT </italic></th><th align="center" rowspan="1" colspan="1"><italic>RPS9 </italic></th><th align="center" rowspan="1" colspan="1"><italic>RPS15A </italic></th><th align="center" rowspan="1" colspan="1"><italic>HMBS </italic></th><th align="center" rowspan="1" colspan="1"><italic>B2M </italic></th><th align="center" rowspan="1" colspan="1"><italic>HPRT1 </italic></th></tr></thead><tbody><tr><td align="left" rowspan="1" colspan="1"><italic>RPL4 </italic></td><td align="center" rowspan="1" colspan="1"><italic>R </italic></td><td align="center" rowspan="1" colspan="1">0.893</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"></td><td align="center" rowspan="1" colspan="1"><italic>P</italic> value</td><td align="center" rowspan="1" colspan="1">0.001</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPS23 </italic></td><td align="center" rowspan="1" colspan="1"><italic>R </italic></td><td align="center" rowspan="1" colspan="1">0.841</td><td align="center" rowspan="1" colspan="1">0.800</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"></td><td align="center" rowspan="1" colspan="1"><italic>P</italic> value</td><td align="center" rowspan="1" colspan="1">0.005</td><td align="center" rowspan="1" colspan="1">0.010</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>GTP </italic></td><td align="center" rowspan="1" colspan="1"><italic>R </italic></td><td align="center" rowspan="1" colspan="1">0.790</td><td align="center" rowspan="1" colspan="1">0.592</td><td align="center" rowspan="1" colspan="1">0.718</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"></td><td align="center" rowspan="1" colspan="1"><italic>P</italic> value</td><td align="center" rowspan="1" colspan="1">0.011</td><td align="center" rowspan="1" colspan="1">0.094</td><td align="center" rowspan="1" colspan="1">0.029</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>UXT </italic></td><td align="center" rowspan="1" colspan="1"><italic>R </italic></td><td align="center" rowspan="1" colspan="1">0.776</td><td align="center" rowspan="1" colspan="1">0.426</td><td align="center" rowspan="1" colspan="1">0.519</td><td align="center" rowspan="1" colspan="1">0.793</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"></td><td align="center" rowspan="1" colspan="1"><italic>P</italic> value</td><td align="center" rowspan="1" colspan="1">0.014</td><td align="center" rowspan="1" colspan="1">0.251</td><td align="center" rowspan="1" colspan="1">0.151</td><td align="center" rowspan="1" colspan="1">0.011</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPS9 </italic></td><td align="center" rowspan="1" colspan="1"><italic>R </italic></td><td align="center" rowspan="1" colspan="1">0.712</td><td align="center" rowspan="1" colspan="1">0.608</td><td align="center" rowspan="1" colspan="1">0.766</td><td align="center" rowspan="1" colspan="1">0.351</td><td align="center" rowspan="1" colspan="1">0.464</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"></td><td align="center" rowspan="1" colspan="1"><italic>P</italic> value</td><td align="center" rowspan="1" colspan="1">0.032</td><td align="center" rowspan="1" colspan="1">0.082</td><td align="center" rowspan="1" colspan="1">0.016</td><td align="center" rowspan="1" colspan="1">0.355</td><td align="center" rowspan="1" colspan="1">0.207</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>RPS15A </italic></td><td align="center" rowspan="1" colspan="1"><italic>R </italic></td><td align="center" rowspan="1" colspan="1">0.507</td><td align="center" rowspan="1" colspan="1">0.250</td><td align="center" rowspan="1" colspan="1">0.592</td><td align="center" rowspan="1" colspan="1">0.788</td><td align="center" rowspan="1" colspan="1">0.634</td><td align="center" rowspan="1" colspan="1">0.363</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"></td><td align="center" rowspan="1" colspan="1"><italic>P</italic> value</td><td align="center" rowspan="1" colspan="1">0.163</td><td align="center" rowspan="1" colspan="1">0.518</td><td align="center" rowspan="1" colspan="1">0.094</td><td align="center" rowspan="1" colspan="1">0.012</td><td align="center" rowspan="1" colspan="1">0.067</td><td align="center" rowspan="1" colspan="1">0.337</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>HMBS </italic></td><td align="center" rowspan="1" colspan="1"><italic>R </italic></td><td align="center" rowspan="1" colspan="1">0.658</td><td align="center" rowspan="1" colspan="1">0.630</td><td align="center" rowspan="1" colspan="1">0.775</td><td align="center" rowspan="1" colspan="1">0.791</td><td align="center" rowspan="1" colspan="1">0.413</td><td align="center" rowspan="1" colspan="1">0.430</td><td align="center" rowspan="1" colspan="1">0.812</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"></td><td align="center" rowspan="1" colspan="1"><italic>P</italic> value</td><td align="center" rowspan="1" colspan="1">0.054</td><td align="center" rowspan="1" colspan="1">0.069</td><td align="center" rowspan="1" colspan="1">0.014</td><td align="center" rowspan="1" colspan="1">0.011</td><td align="center" rowspan="1" colspan="1">0.269</td><td align="center" rowspan="1" colspan="1">0.248</td><td align="center" rowspan="1" colspan="1">0.008</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>B2M </italic></td><td align="center" rowspan="1" colspan="1"><italic>R </italic></td><td align="center" rowspan="1" colspan="1">0.206</td><td align="center" rowspan="1" colspan="1">0.056</td><td align="center" rowspan="1" colspan="1">0.072</td><td align="center" rowspan="1" colspan="1">0.179</td><td align="center" rowspan="1" colspan="1">0.407</td><td align="center" rowspan="1" colspan="1">−0.042</td><td align="center" rowspan="1" colspan="1">0.384</td><td align="center" rowspan="1" colspan="1">0.168</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"></td><td align="center" rowspan="1" colspan="1"><italic>P</italic> value</td><td align="center" rowspan="1" colspan="1">0.593</td><td align="center" rowspan="1" colspan="1">0.885</td><td align="center" rowspan="1" colspan="1">0.855</td><td align="center" rowspan="1" colspan="1">0.646</td><td align="center" rowspan="1" colspan="1">0.277</td><td align="center" rowspan="1" colspan="1">0.915</td><td align="center" rowspan="1" colspan="1">0.308</td><td align="center" rowspan="1" colspan="1">0.666</td><td align="center" rowspan="1" colspan="1">—</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"><italic>HPRT1 </italic></td><td align="center" rowspan="1" colspan="1"><italic>R </italic></td><td align="center" rowspan="1" colspan="1">0.382</td><td align="center" rowspan="1" colspan="1">0.305</td><td align="center" rowspan="1" colspan="1">0.273</td><td align="center" rowspan="1" colspan="1">0.639</td><td align="center" rowspan="1" colspan="1">0.368</td><td align="center" rowspan="1" colspan="1">0.146</td><td align="center" rowspan="1" colspan="1">0.676</td><td align="center" rowspan="1" colspan="1">0.743</td><td align="center" rowspan="1" colspan="1">−0.013</td><td align="center" rowspan="1" colspan="1">—</td></tr><tr><td align="left" rowspan="1" colspan="1"></td><td align="center" rowspan="1" colspan="1"><italic>P</italic> value</td><td align="center" rowspan="1" colspan="1">0.312</td><td align="center" rowspan="1" colspan="1">0.423</td><td align="center" rowspan="1" colspan="1">0.478</td><td align="center" rowspan="1" colspan="1">0.064</td><td align="center" rowspan="1" colspan="1">0.329</td><td align="center" rowspan="1" colspan="1">0.708</td><td align="center" rowspan="1" colspan="1">0.045</td><td align="center" rowspan="1" colspan="1">0.022</td><td align="center" rowspan="1" colspan="1">0.977</td><td align="center" rowspan="1" colspan="1"><italic>— </italic></td></tr><tr><td align="left" rowspan="3" colspan="1">BestKeeper Index</td><td align="center" rowspan="1" colspan="1"><italic>R </italic></td><td align="center" rowspan="1" colspan="1">0.804</td><td align="center" rowspan="1" colspan="1">0.679</td><td align="center" rowspan="1" colspan="1">0.840</td><td align="center" rowspan="1" colspan="1">0.503</td><td align="center" rowspan="1" colspan="1">0.565</td><td align="center" rowspan="1" colspan="1">0.983</td><td align="center" rowspan="1" colspan="1">0.497</td><td align="center" rowspan="1" colspan="1">0.567</td><td align="center" rowspan="1" colspan="1">0.035</td><td align="center" rowspan="1" colspan="1">0.273</td></tr><tr><td align="center" rowspan="1" colspan="1"><italic>P</italic> value</td><td align="center" rowspan="1" colspan="1">0.009</td><td align="center" rowspan="1" colspan="1">0.044</td><td align="center" rowspan="1" colspan="1">0.005</td><td align="center" rowspan="1" colspan="1">0.167</td><td align="center" rowspan="1" colspan="1">0.113</td><td align="center" rowspan="1" colspan="1">0.001</td><td align="center" rowspan="1" colspan="1">0.172</td><td align="center" rowspan="1" colspan="1">0.112</td><td align="center" rowspan="1" colspan="1">0.931</td><td align="center" rowspan="1" colspan="1">0.478</td></tr><tr><td align="center" rowspan="1" colspan="1"><italic>r</italic><sup>2</sup></td><td align="center" rowspan="1" colspan="1">0.646</td><td align="center" rowspan="1" colspan="1">0.461</td><td align="center" rowspan="1" colspan="1">0.706</td><td align="center" rowspan="1" colspan="1">0.253</td><td align="center" rowspan="1" colspan="1">0.319</td><td align="center" rowspan="1" colspan="1">0.966</td><td align="center" rowspan="1" colspan="1">0.247</td><td align="center" rowspan="1" colspan="1">0.321</td><td align="center" rowspan="1" colspan="1">0.001</td><td align="center" rowspan="1" colspan="1">0.075</td></tr></tbody></table><table-wrap-foot><fn><p><italic>R</italic>: Pearson correlation coefficient; <italic>r</italic><sup>2</sup>: coefficient of determination.</p></fn></table-wrap-foot></table-wrap></floats-group></pmc><affiliations><list><country><li>Inde</li></country></list><tree><country name="Inde"><noRegion><name sortKey="Kapila, Neha" sort="Kapila, Neha" uniqKey="Kapila N" first="Neha" last="Kapila">Neha Kapila</name></noRegion><name sortKey="Jerath, Tanushri" sort="Jerath, Tanushri" uniqKey="Jerath T" first="Tanushri" last="Jerath">Tanushri Jerath</name><name sortKey="Kapila, Neha" sort="Kapila, Neha" uniqKey="Kapila N" first="Neha" last="Kapila">Neha Kapila</name><name sortKey="Kishore, Amit" sort="Kishore, Amit" uniqKey="Kishore A" first="Amit" last="Kishore">Amit Kishore</name><name sortKey="Kumar, Pawan" sort="Kumar, Pawan" uniqKey="Kumar P" first="Pawan" last="Kumar">Pawan Kumar</name><name sortKey="Mohanty, A K" sort="Mohanty, A K" uniqKey="Mohanty A" first="A. K." last="Mohanty">A. K. Mohanty</name><name sortKey="Mukesh, M" sort="Mukesh, M" uniqKey="Mukesh M" first="M." last="Mukesh">M. Mukesh</name><name sortKey="Sharma, Ankita" sort="Sharma, Ankita" uniqKey="Sharma A" first="Ankita" last="Sharma">Ankita Sharma</name><name sortKey="Sodhi, Monika" sort="Sodhi, Monika" uniqKey="Sodhi M" first="Monika" last="Sodhi">Monika Sodhi</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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