Xanthium strumarium Fruit Extract Inhibits ATG4B and Diminishes the Proliferation and Metastatic Characteristics of Colorectal Cancer Cells
Identifieur interne : 000833 ( Pmc/Curation ); précédent : 000832; suivant : 000834Xanthium strumarium Fruit Extract Inhibits ATG4B and Diminishes the Proliferation and Metastatic Characteristics of Colorectal Cancer Cells
Auteurs : Hsueh-Wei Chang [Taïwan] ; Pei-Feng Liu ; Wei-Lun Tsai ; Wan-Hsiang Hu ; Yu-Chang Hu ; Hsiu-Chen Yang ; Wei-Yu Lin ; Jing-Ru Weng [Taïwan] ; Chih-Wen Shu [Taïwan]Source :
- Toxins [ 2072-6651 ] ; 2019.
Abstract
Autophagy is an evolutionarily conserved pathway to degrade damaged proteins and organelles for subsequent recycling in cells during times of nutrient deprivation. This process plays an important role in tumor development and progression, allowing cancer cells to survive in nutrient-poor environments. The plant kingdom provides a powerful source for new drug development to treat cancer. Several plant extracts induce autophagy in cancer cells. However, little is known about the role of plant extracts in autophagy inhibition, particularly autophagy-related (ATG) proteins. In this study, we employed S-tagged gamma-aminobutyric acid receptor associated protein like 2 (GABARAPL2) as a reporter to screen 48 plant extracts for their effects on the activity of autophagy protease ATG4B.
Url:
DOI: 10.3390/toxins11060313
PubMed: 31159487
PubMed Central: 6628400
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en"><italic>Xanthium strumarium</italic> Fruit Extract Inhibits ATG4B and Diminishes the Proliferation and Metastatic Characteristics of Colorectal Cancer Cells</title><author><name sortKey="Chang, Hsueh Wei" sort="Chang, Hsueh Wei" uniqKey="Chang H" first="Hsueh-Wei" last="Chang">Hsueh-Wei Chang</name><affiliation><nlm:aff id="af1-toxins-11-00313">Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung 80708, Taiwan;<email>changhw@kmu.edu.tw</email></nlm:aff></affiliation><affiliation wicri:level="1"><nlm:aff id="af2-toxins-11-00313">Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung 80708, Taiwan</nlm:aff><country xml:lang="fr">Taïwan</country><wicri:regionArea>Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung 80708</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="af3-toxins-11-00313">Drug Development and Value Creation Research Center, Kaohsiung Medical University, Kaohsiung 80708, Taiwan</nlm:aff><country xml:lang="fr">Taïwan</country><wicri:regionArea>Drug Development and Value Creation Research Center, Kaohsiung Medical University, Kaohsiung 80708</wicri:regionArea></affiliation></author><author><name sortKey="Liu, Pei Feng" sort="Liu, Pei Feng" uniqKey="Liu P" first="Pei-Feng" last="Liu">Pei-Feng Liu</name><affiliation><nlm:aff id="af4-toxins-11-00313">Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan;<email>pfliu@vghks.gov.tw</email> (P.-F.L.);<email>mini16610@gmail.com</email> (H.-C.Y.)</nlm:aff></affiliation></author><author><name sortKey="Tsai, Wei Lun" sort="Tsai, Wei Lun" uniqKey="Tsai W" first="Wei-Lun" last="Tsai">Wei-Lun Tsai</name><affiliation><nlm:aff id="af5-toxins-11-00313">Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Veterans General Hospital, Kaohsiung 813, Taiwan;<email>tsaiwl@yahoo.com.tw</email></nlm:aff></affiliation></author><author><name sortKey="Hu, Wan Hsiang" sort="Hu, Wan Hsiang" uniqKey="Hu W" first="Wan-Hsiang" last="Hu">Wan-Hsiang Hu</name><affiliation><nlm:aff id="af6-toxins-11-00313">Department of Colorectal Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung 83341, Taiwan;<email>gary.hu0805@msa.hinet.net</email></nlm:aff></affiliation></author><author><name sortKey="Hu, Yu Chang" sort="Hu, Yu Chang" uniqKey="Hu Y" first="Yu-Chang" last="Hu">Yu-Chang Hu</name><affiliation><nlm:aff id="af7-toxins-11-00313">Department of Radiation Oncology, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan;<email>ychu81@gmail.com</email></nlm:aff></affiliation></author><author><name sortKey="Yang, Hsiu Chen" sort="Yang, Hsiu Chen" uniqKey="Yang H" first="Hsiu-Chen" last="Yang">Hsiu-Chen Yang</name><affiliation><nlm:aff id="af4-toxins-11-00313">Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan;<email>pfliu@vghks.gov.tw</email> (P.-F.L.);<email>mini16610@gmail.com</email> (H.-C.Y.)</nlm:aff></affiliation></author><author><name sortKey="Lin, Wei Yu" sort="Lin, Wei Yu" uniqKey="Lin W" first="Wei-Yu" last="Lin">Wei-Yu Lin</name><affiliation><nlm:aff id="af8-toxins-11-00313">Department of Pharmacy, Kinmen Hospital, Kinmen 89142, Taiwan;<email>u8557006@gmail.com</email></nlm:aff></affiliation></author><author><name sortKey="Weng, Jing Ru" sort="Weng, Jing Ru" uniqKey="Weng J" first="Jing-Ru" last="Weng">Jing-Ru Weng</name><affiliation wicri:level="1"><nlm:aff id="af9-toxins-11-00313">Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 80424, Taiwan</nlm:aff><country xml:lang="fr">Taïwan</country><wicri:regionArea>Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 80424</wicri:regionArea></affiliation></author><author><name sortKey="Shu, Chih Wen" sort="Shu, Chih Wen" uniqKey="Shu C" first="Chih-Wen" last="Shu">Chih-Wen Shu</name><affiliation wicri:level="1"><nlm:aff id="af10-toxins-11-00313">School of Medicine for International Students, I-Shou University, Kaohsiung 82445, Taiwan</nlm:aff><country xml:lang="fr">Taïwan</country><wicri:regionArea>School of Medicine for International Students, I-Shou University, Kaohsiung 82445</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">31159487</idno><idno type="pmc">6628400</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6628400</idno><idno type="RBID">PMC:6628400</idno><idno type="doi">10.3390/toxins11060313</idno><date when="2019">2019</date><idno type="wicri:Area/Pmc/Corpus">000833</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000833</idno><idno type="wicri:Area/Pmc/Curation">000833</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000833</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main"><italic>Xanthium strumarium</italic> Fruit Extract Inhibits ATG4B and Diminishes the Proliferation and Metastatic Characteristics of Colorectal Cancer Cells</title><author><name sortKey="Chang, Hsueh Wei" sort="Chang, Hsueh Wei" uniqKey="Chang H" first="Hsueh-Wei" last="Chang">Hsueh-Wei Chang</name><affiliation><nlm:aff id="af1-toxins-11-00313">Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung 80708, Taiwan;<email>changhw@kmu.edu.tw</email></nlm:aff></affiliation><affiliation wicri:level="1"><nlm:aff id="af2-toxins-11-00313">Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung 80708, Taiwan</nlm:aff><country xml:lang="fr">Taïwan</country><wicri:regionArea>Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung 80708</wicri:regionArea></affiliation><affiliation wicri:level="1"><nlm:aff id="af3-toxins-11-00313">Drug Development and Value Creation Research Center, Kaohsiung Medical University, Kaohsiung 80708, Taiwan</nlm:aff><country xml:lang="fr">Taïwan</country><wicri:regionArea>Drug Development and Value Creation Research Center, Kaohsiung Medical University, Kaohsiung 80708</wicri:regionArea></affiliation></author><author><name sortKey="Liu, Pei Feng" sort="Liu, Pei Feng" uniqKey="Liu P" first="Pei-Feng" last="Liu">Pei-Feng Liu</name><affiliation><nlm:aff id="af4-toxins-11-00313">Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan;<email>pfliu@vghks.gov.tw</email> (P.-F.L.);<email>mini16610@gmail.com</email> (H.-C.Y.)</nlm:aff></affiliation></author><author><name sortKey="Tsai, Wei Lun" sort="Tsai, Wei Lun" uniqKey="Tsai W" first="Wei-Lun" last="Tsai">Wei-Lun Tsai</name><affiliation><nlm:aff id="af5-toxins-11-00313">Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Veterans General Hospital, Kaohsiung 813, Taiwan;<email>tsaiwl@yahoo.com.tw</email></nlm:aff></affiliation></author><author><name sortKey="Hu, Wan Hsiang" sort="Hu, Wan Hsiang" uniqKey="Hu W" first="Wan-Hsiang" last="Hu">Wan-Hsiang Hu</name><affiliation><nlm:aff id="af6-toxins-11-00313">Department of Colorectal Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung 83341, Taiwan;<email>gary.hu0805@msa.hinet.net</email></nlm:aff></affiliation></author><author><name sortKey="Hu, Yu Chang" sort="Hu, Yu Chang" uniqKey="Hu Y" first="Yu-Chang" last="Hu">Yu-Chang Hu</name><affiliation><nlm:aff id="af7-toxins-11-00313">Department of Radiation Oncology, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan;<email>ychu81@gmail.com</email></nlm:aff></affiliation></author><author><name sortKey="Yang, Hsiu Chen" sort="Yang, Hsiu Chen" uniqKey="Yang H" first="Hsiu-Chen" last="Yang">Hsiu-Chen Yang</name><affiliation><nlm:aff id="af4-toxins-11-00313">Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan;<email>pfliu@vghks.gov.tw</email> (P.-F.L.);<email>mini16610@gmail.com</email> (H.-C.Y.)</nlm:aff></affiliation></author><author><name sortKey="Lin, Wei Yu" sort="Lin, Wei Yu" uniqKey="Lin W" first="Wei-Yu" last="Lin">Wei-Yu Lin</name><affiliation><nlm:aff id="af8-toxins-11-00313">Department of Pharmacy, Kinmen Hospital, Kinmen 89142, Taiwan;<email>u8557006@gmail.com</email></nlm:aff></affiliation></author><author><name sortKey="Weng, Jing Ru" sort="Weng, Jing Ru" uniqKey="Weng J" first="Jing-Ru" last="Weng">Jing-Ru Weng</name><affiliation wicri:level="1"><nlm:aff id="af9-toxins-11-00313">Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 80424, Taiwan</nlm:aff><country xml:lang="fr">Taïwan</country><wicri:regionArea>Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 80424</wicri:regionArea></affiliation></author><author><name sortKey="Shu, Chih Wen" sort="Shu, Chih Wen" uniqKey="Shu C" first="Chih-Wen" last="Shu">Chih-Wen Shu</name><affiliation wicri:level="1"><nlm:aff id="af10-toxins-11-00313">School of Medicine for International Students, I-Shou University, Kaohsiung 82445, Taiwan</nlm:aff><country xml:lang="fr">Taïwan</country><wicri:regionArea>School of Medicine for International Students, I-Shou University, Kaohsiung 82445</wicri:regionArea></affiliation></author></analytic><series><title level="j">Toxins</title><idno type="eISSN">2072-6651</idno><imprint><date when="2019">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Autophagy is an evolutionarily conserved pathway to degrade damaged proteins and organelles for subsequent recycling in cells during times of nutrient deprivation. This process plays an important role in tumor development and progression, allowing cancer cells to survive in nutrient-poor environments. The plant kingdom provides a powerful source for new drug development to treat cancer. Several plant extracts induce autophagy in cancer cells. However, little is known about the role of plant extracts in autophagy inhibition, particularly autophagy-related (ATG) proteins. In this study, we employed S-tagged gamma-aminobutyric acid receptor associated protein like 2 (GABARAPL2) as a reporter to screen 48 plant extracts for their effects on the activity of autophagy protease ATG4B. <italic>Xanthium strumarium</italic> and <italic>Tribulus terrestris</italic> fruit extracts were validated as potential ATG4B inhibitors by another reporter substrate MAP1LC3B-PLA<sub>2</sub>. The inhibitory effects of the extracts on cellular ATG4B and autophagic flux were further confirmed. Moreover, the plant extracts significantly reduced colorectal cancer cell viability and sensitized cancer cells to starvation conditions. The fruit extract of <italic>X. strumarium</italic> consistently diminished cancer cell migration and invasion. Taken together, the results showed that the fruit of <italic>X. strumarium</italic> may have an active ingredient to inhibit ATG4B and suppress the proliferation and metastatic characteristics of colorectal cancer cells. </p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Amaravadi, R" uniqKey="Amaravadi R">R. Amaravadi</name></author><author><name sortKey="Kimmelman, A C" uniqKey="Kimmelman A">A.C. Kimmelman</name></author><author><name sortKey="White, E" uniqKey="White E">E. White</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Takamura, A" uniqKey="Takamura A">A. Takamura</name></author><author><name sortKey="Komatsu, M" uniqKey="Komatsu M">M. 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Toxins (Basel)</journal-id><journal-id journal-id-type="iso-abbrev">Toxins (Basel)</journal-id><journal-id journal-id-type="publisher-id">toxins</journal-id><journal-title-group><journal-title>Toxins</journal-title></journal-title-group><issn pub-type="epub">2072-6651</issn><publisher><publisher-name>MDPI</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">31159487</article-id><article-id pub-id-type="pmc">6628400</article-id><article-id pub-id-type="doi">10.3390/toxins11060313</article-id><article-id pub-id-type="publisher-id">toxins-11-00313</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title><italic>Xanthium strumarium</italic> Fruit Extract Inhibits ATG4B and Diminishes the Proliferation and Metastatic Characteristics of Colorectal Cancer Cells</article-title></title-group><contrib-group><contrib contrib-type="author"><contrib-id contrib-id-type="orcid" authenticated="true">https://orcid.org/0000-0003-0068-2366</contrib-id><name><surname>Chang</surname><given-names>Hsueh-Wei</given-names></name><xref ref-type="aff" rid="af1-toxins-11-00313">1</xref><xref ref-type="aff" rid="af2-toxins-11-00313">2</xref><xref ref-type="aff" rid="af3-toxins-11-00313">3</xref><xref ref-type="author-notes" rid="fn1-toxins-11-00313">†</xref></contrib><contrib contrib-type="author"><name><surname>Liu</surname><given-names>Pei-Feng</given-names></name><xref ref-type="aff" rid="af4-toxins-11-00313">4</xref><xref ref-type="author-notes" rid="fn1-toxins-11-00313">†</xref></contrib><contrib contrib-type="author"><name><surname>Tsai</surname><given-names>Wei-Lun</given-names></name><xref ref-type="aff" rid="af5-toxins-11-00313">5</xref></contrib><contrib contrib-type="author"><name><surname>Hu</surname><given-names>Wan-Hsiang</given-names></name><xref ref-type="aff" rid="af6-toxins-11-00313">6</xref></contrib><contrib contrib-type="author"><name><surname>Hu</surname><given-names>Yu-Chang</given-names></name><xref ref-type="aff" rid="af7-toxins-11-00313">7</xref></contrib><contrib contrib-type="author"><name><surname>Yang</surname><given-names>Hsiu-Chen</given-names></name><xref ref-type="aff" rid="af4-toxins-11-00313">4</xref></contrib><contrib contrib-type="author"><name><surname>Lin</surname><given-names>Wei-Yu</given-names></name><xref ref-type="aff" rid="af8-toxins-11-00313">8</xref></contrib><contrib contrib-type="author"><name><surname>Weng</surname><given-names>Jing-Ru</given-names></name><xref ref-type="aff" rid="af9-toxins-11-00313">9</xref><xref rid="c1-toxins-11-00313" ref-type="corresp">*</xref></contrib><contrib contrib-type="author"><contrib-id contrib-id-type="orcid" authenticated="true">https://orcid.org/0000-0002-7774-0002</contrib-id><name><surname>Shu</surname><given-names>Chih-Wen</given-names></name><xref ref-type="aff" rid="af10-toxins-11-00313">10</xref><xref rid="c1-toxins-11-00313" ref-type="corresp">*</xref></contrib></contrib-group><aff id="af1-toxins-11-00313"><label>1</label>Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung 80708, Taiwan;<email>changhw@kmu.edu.tw</email></aff><aff id="af2-toxins-11-00313"><label>2</label>Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung 80708, Taiwan</aff><aff id="af3-toxins-11-00313"><label>3</label>Drug Development and Value Creation Research Center, Kaohsiung Medical University, Kaohsiung 80708, Taiwan</aff><aff id="af4-toxins-11-00313"><label>4</label>Department of Medical Education and Research, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan;<email>pfliu@vghks.gov.tw</email> (P.-F.L.);<email>mini16610@gmail.com</email> (H.-C.Y.)</aff><aff id="af5-toxins-11-00313"><label>5</label>Division of Gastroenterology, Department of Internal Medicine, Kaohsiung Veterans General Hospital, Kaohsiung 813, Taiwan;<email>tsaiwl@yahoo.com.tw</email></aff><aff id="af6-toxins-11-00313"><label>6</label>Department of Colorectal Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung 83341, Taiwan;<email>gary.hu0805@msa.hinet.net</email></aff><aff id="af7-toxins-11-00313"><label>7</label>Department of Radiation Oncology, Kaohsiung Veterans General Hospital, Kaohsiung 81362, Taiwan;<email>ychu81@gmail.com</email></aff><aff id="af8-toxins-11-00313"><label>8</label>Department of Pharmacy, Kinmen Hospital, Kinmen 89142, Taiwan;<email>u8557006@gmail.com</email></aff><aff id="af9-toxins-11-00313"><label>9</label>Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung 80424, Taiwan</aff><aff id="af10-toxins-11-00313"><label>10</label>School of Medicine for International Students, I-Shou University, Kaohsiung 82445, Taiwan</aff><author-notes><corresp id="c1-toxins-11-00313"><label>*</label>Correspondence: <email>jrweng@mail.nsysu.edu.tw</email> (J.-R.W.); <email>cwshu@isu.edu.tw</email> (C.-W.S.); Tel.: +886-7525-2000 (J.-R.W.); +886-7615-1100 (C.-W.S.)</corresp><fn id="fn1-toxins-11-00313"><label>†</label><p>These two authors contribute equally to this work.</p></fn></author-notes><pub-date pub-type="epub"><day>02</day><month>6</month><year>2019</year></pub-date><pub-date pub-type="collection"><month>6</month><year>2019</year></pub-date><volume>11</volume><issue>6</issue><elocation-id>313</elocation-id><history><date date-type="received"><day>06</day><month>5</month><year>2019</year></date><date date-type="accepted"><day>30</day><month>5</month><year>2019</year></date></history><permissions><copyright-statement>© 2019 by the authors.</copyright-statement><copyright-year>2019</copyright-year><license license-type="open-access"><license-p>Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">http://creativecommons.org/licenses/by/4.0/</ext-link>).</license-p></license></permissions><abstract><p>Autophagy is an evolutionarily conserved pathway to degrade damaged proteins and organelles for subsequent recycling in cells during times of nutrient deprivation. This process plays an important role in tumor development and progression, allowing cancer cells to survive in nutrient-poor environments. The plant kingdom provides a powerful source for new drug development to treat cancer. Several plant extracts induce autophagy in cancer cells. However, little is known about the role of plant extracts in autophagy inhibition, particularly autophagy-related (ATG) proteins. In this study, we employed S-tagged gamma-aminobutyric acid receptor associated protein like 2 (GABARAPL2) as a reporter to screen 48 plant extracts for their effects on the activity of autophagy protease ATG4B. <italic>Xanthium strumarium</italic> and <italic>Tribulus terrestris</italic> fruit extracts were validated as potential ATG4B inhibitors by another reporter substrate MAP1LC3B-PLA<sub>2</sub>. The inhibitory effects of the extracts on cellular ATG4B and autophagic flux were further confirmed. Moreover, the plant extracts significantly reduced colorectal cancer cell viability and sensitized cancer cells to starvation conditions. The fruit extract of <italic>X. strumarium</italic> consistently diminished cancer cell migration and invasion. Taken together, the results showed that the fruit of <italic>X. strumarium</italic> may have an active ingredient to inhibit ATG4B and suppress the proliferation and metastatic characteristics of colorectal cancer cells. </p></abstract><kwd-group><kwd>medicinal plant</kwd><kwd>autophagy</kwd><kwd>ATG4B</kwd><kwd>colorectal cancer</kwd></kwd-group></article-meta><notes notes-type="Key Contribution"><title>Key Contribution</title><p><italic>X. strumarium</italic> fruit extract inhibits ATG4B and autophagy. <italic>X. strumarium</italic> fruit extract reduces cell proliferation and metastatic characteristics of colorectal cancer cells.</p></notes></front><floats-group><fig id="toxins-11-00313-f001" orientation="portrait" position="float"><label>Figure 1</label><caption><p>Screening for ATG4B inhibitors with medicinal plants using ATG4B cleavage assays. (<bold>A</bold>) Schematic diagram for the biochemical reaction to measure ATG4B proteolytic activity. Briefly, the substrate GABARAPL2 fused with the N-terminal C-myc tag and C-terminal S-tag. If extracts inhibited ATG4B, both full-length C-myc-tagged GABARAP2 (upper band) and a short peptide of S-tag can be detected by immunoblotting. (<bold>B</bold>) Recombinant ATG4B (5 nM) was mixed with the GABARAPL2 fusion protein (1 μM) in a reaction buffer containing 50 mM Tris-base, pH 8.0, 150 mM NaCl, and 1 mM DTT with 10-fold diluted formosan plant extracts (10 or 100 μg/mL). The retained S-tag and full-length C-myc tag (upper band) were determined by immunoblotting. The reactions without (+) and with (−) recombinant ATG4B were used as positive and negative controls of inhibition, respectively. (<bold>C</bold>) The protein levels of S-tag and (<bold>D</bold>) full-length C-myc-tagged GABARAPL2 were quantitated by ImageJ and normalized by positive control (+) as 100% inhibition of ATG4B in each blot.</p></caption><graphic xlink:href="toxins-11-00313-g001"></graphic></fig><fig id="toxins-11-00313-f002" orientation="portrait" position="float"><label>Figure 2</label><caption><p>Hits validation of ATG4B inhibitors using the LC3B-PLA<sub>2</sub> biochemical assay. Four hits obtained from the ATG4B cleavage assay—(<bold>A</bold>) XS-(fr)-M, (<bold>B</bold>) XS-(fr)-C, (<bold>C</bold>) TT-(fr)-A and (<bold>D</bold>) TT-(fr)-M—were reconfirmed using the MAP1LC3B-PLA<sub>2</sub> biochemical assay. Medicinal plant extracts were mixed with recombinant ATG4B (0.5 nM) in the presence of 500 nM of the reporter MAP1LC3B-PLA<sub>2</sub>, another substrate for ATG4B, in a reaction buffer to determine the inhibitory effects of extracts on ATG4B activity. The fluorescence intensity was repeatedly measured for 150 min with excitation and emission wavelength of 485 and 530, respectively. The results are expressed as the mean ± SEM from three individual experiments.</p></caption><graphic xlink:href="toxins-11-00313-g002"></graphic></fig><fig id="toxins-11-00313-f003" orientation="portrait" position="float"><label>Figure 3</label><caption><p>Effects of medicinal plants on cellular ATG4B and autophagic flux in human cells. (<bold>A</bold>) HEK293T cells were transfected with ATG4-cleavable luciferase and then were treated with plant extracts, including XS-(fr)-M, XS-(fr)-C, TT-(fr)-A, and TT-(fr)-M, for 6 h. The cells were mixed with One-Glo to measure luciferase activity. Increased luciferase activity indicated ATG4B inhibition, as described in the method. (<bold>B</bold>) To measure autophagic flux, HCT116 cells were treated with medicinal plant extracts in a cultured medium in the presence or absence of the autophagy inhibitor bafilomycin A1(B, 100 nM) for 3 h. The cells were then harvested for immunoblotting, using antibodies against MAP1LC3B and TUBA to determine MAP1LC3B-II accumulation to reflect autophagic flux. (<bold>C</bold>) The quantitative results for panel B were expressed as the means ± SEM from three independent experiments. The results are expressed as the mean ± SEM from 3 individual experiments. * <italic>p</italic> < 0.05; ** <italic>p</italic> < 0.01; *** <italic>p</italic> < 0.001 vs. cells treated with DMSO (0 or Control).</p></caption><graphic xlink:href="toxins-11-00313-g003"></graphic></fig><fig id="toxins-11-00313-f004" orientation="portrait" position="float"><label>Figure 4</label><caption><p>Effects of compounds on cell viability in colorectal cancer cells. Human colorectal cancer HCT116 and DLD-1 cells were cultured and treated with 10-fold diluted (10 or 100 μg/mL) extracts (<bold>A</bold>) XS-(fr)-M, (<bold>B</bold>) XS-(fr)-C, (<bold>C</bold>) TT-(fr)-A, (<bold>D</bold>) TT-(fr)-M in cultured medium (DMEM) or autophagy-inducing medium Earle’s Balanced Salt Solution (EBSS) for 24 h. The CellTiter-Glo reagent was added to cells, followed by measurement of the cellular ATP level to reflect cell viability. The results are expressed as the mean ± SEM from three individual experiments. * <italic>p</italic> < 0.05; ** <italic>p</italic> < 0.01; *** <italic>p</italic> < 0.001 for cells in starvation medium EBSS vs. cells with the same treatment in regular medium (Control).</p></caption><graphic xlink:href="toxins-11-00313-g004"></graphic></fig><fig id="toxins-11-00313-f005" orientation="portrait" position="float"><label>Figure 5</label><caption><p>Effects of plant extracts on the migration of human colorectal cancer cells. Human colorectal cancer HCT116 (<bold>A</bold>,<bold>C</bold>) or DLD-1 (<bold>B</bold>,<bold>D</bold>) cells were seeded in culture inserts overnight and then were treated with plant extracts XS-fr-C (<bold>A</bold>,<bold>B</bold>) or TT-fr-M (<bold>C</bold>,<bold>D</bold>) for 16 h. The wounds were imaged and quantified by ImageJ software. The effects of plant extracts on cell migration were analyzed by Prism 5.0. The results are expressed as the mean ± SEM from three individual experiments. * <italic>p</italic> < 0.05; ** <italic>p</italic> < 0.01; *** <italic>p</italic> < 0.001 vs. cells treated with DMSO (0).</p></caption><graphic xlink:href="toxins-11-00313-g005"></graphic></fig><fig id="toxins-11-00313-f006" orientation="portrait" position="float"><label>Figure 6</label><caption><p>Effects of plant extracts on cell invasion in colorectal cancer cells. Human colorectal cancer HCT116 (<bold>A</bold>) or DLD-1 (<bold>C</bold>) cells were seeded on the top chamber of Matrigel-coated filters with low-FBS medium in the presence or absence (-) of plant extracts (<bold>A</bold>) XS-fr-C or (<bold>C</bold>) TT-fr-M. After invasion for 24 h, the filters were removed for fixation and stained with crystal violet. Scale Bars: 50 µm. The number of invaded cells was imaged with a microscope. A representative image is shown, and quantitative results (<bold>B</bold>,<bold>D</bold>) were expressed as the means ± SEM from three independent experiments. The results are expressed as the mean ± SEM from three individual experiments. * <italic>p</italic> < 0.05; ** <italic>p</italic> < 0.01; *** <italic>p</italic> < 0.001 vs. cells treated with DMSO (0). (<bold>E</bold>) HCT116 cells were cultured on NanoCulture plates for tumor sphere formation and treated with XS-fr-C or TT-fr-M for 48 h. The viable (green) and dead (red) cells were stained by a LIVE/DEAD staining kit. The representative images are shown. (<bold>F</bold>) The viable and dead cells were quantitated by a florescent reader. Scale Bars: 100 µm.</p></caption><graphic xlink:href="toxins-11-00313-g006"></graphic></fig><table-wrap id="toxins-11-00313-t001" orientation="portrait" position="float"><object-id pub-id-type="pii">toxins-11-00313-t001_Table 1</object-id><label>Table 1</label><caption><p>Plant species tested for potential ATG4B inhibitors.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="center" valign="middle" style="border-top:solid thin;border-bottom:solid thin" rowspan="1" colspan="1">No.</th><th align="center" valign="middle" style="border-top:solid thin;border-bottom:solid thin" rowspan="1" colspan="1">Botanical Name</th><th align="center" valign="middle" style="border-top:solid thin;border-bottom:solid thin" rowspan="1" colspan="1">Extract</th><th align="center" valign="middle" style="border-top:solid thin;border-bottom:solid thin" rowspan="1" colspan="1">Family</th><th align="center" valign="middle" style="border-top:solid thin;border-bottom:solid thin" rowspan="1" colspan="1">Part of Plant</th></tr></thead><tbody><tr><td align="center" valign="middle" rowspan="1" colspan="1">1</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Saurauia tristyla</italic> var. <italic>oldhamii</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">Acetone</td><td align="center" valign="middle" rowspan="1" colspan="1">Rubiaceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">2</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">CHCl<sub>3</sub></td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">3</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">4</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Fraxinus griffithii</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">Acetone</td><td align="center" valign="middle" rowspan="1" colspan="1">Oleaceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">5</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">CHCl<sub>3</sub></td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">6</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">7</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Phytolacca americana</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">Acetone</td><td align="center" valign="middle" rowspan="1" colspan="1">Phytolaccaceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Whole plant</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">8</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">CHCl<sub>3</sub></td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Whole plant</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">9</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Whole plant</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">10</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Elaeocarpus sylvestris</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">Acetone</td><td align="center" valign="middle" rowspan="1" colspan="1">Elaeocarpaceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">11</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">CHCl<sub>3</sub></td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">12</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">13</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Stem</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">14</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Piper kadsura</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1">Piperaceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Stem</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">15</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Itea parviflora</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1">Saxifragaceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Stem</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">16</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Callicarpa formosana</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">Acetone</td><td align="center" valign="middle" rowspan="1" colspan="1">Verbenaceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">17</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">18</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Stem</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">19</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Callicarpa kochiana</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">Acetone</td><td align="center" valign="middle" rowspan="1" colspan="1">Verbenaceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">20</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">CHCl<sub>3</sub></td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">21</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">22</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Stem</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">23</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Stem</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">24</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Ficus septica</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">Acetone</td><td align="center" valign="middle" rowspan="1" colspan="1">Moraceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">25</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">CHCl<sub>3</sub></td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">26</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">27</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Fruit</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">28</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Heartwood</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">29</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Ficus sarmentosa</italic> var. <italic>henryi</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">Acetone</td><td align="center" valign="middle" rowspan="1" colspan="1">Moraceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">30</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">CHCl<sub>3</sub></td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">31</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">32</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Stem</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">33</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Xanthium strumarium</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">CHCl<sub>3</sub></td><td align="center" valign="middle" rowspan="1" colspan="1">Asteraceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Fruit</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">34</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Fruit</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">35</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Tribulus terrestris</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">Acetone</td><td align="center" valign="middle" rowspan="1" colspan="1">Zygophyllaceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Fruit</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">36</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Fruit</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">37</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Cornus officinalis</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">Acetone</td><td align="center" valign="middle" rowspan="1" colspan="1">Cornaceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Whole plant</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">38</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1"></td><td align="center" valign="middle" rowspan="1" colspan="1">Whole plant</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">39</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Alisma orientale</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1">Alismataceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Whole plant</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">40</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Asparagus cochinchinensis</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1">Asparagaceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Root</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">41</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Broussonetia papyrifera</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1">Moraceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">42</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Clausena excavata</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1">Rutaceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">43</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Cinnamomum insulari-montanum</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1">Lauraceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">44</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Lumnitzera racemosa</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1">Combretaceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Leaves</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">45</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Pueraria lobata</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1">Fabaceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Whole plant</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">46</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Sida acuta</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1">Malvaceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Whole plant</td></tr><tr><td align="center" valign="middle" rowspan="1" colspan="1">47</td><td align="center" valign="middle" rowspan="1" colspan="1"><italic>Scrophularia ningpoensis</italic></td><td align="center" valign="middle" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" rowspan="1" colspan="1">Scrophulariaceae</td><td align="center" valign="middle" rowspan="1" colspan="1">Whole plant</td></tr><tr><td align="center" valign="middle" style="border-bottom:solid thin" rowspan="1" colspan="1">48</td><td align="center" valign="middle" style="border-bottom:solid thin" rowspan="1" colspan="1"><italic>Catharanthus roseus</italic></td><td align="center" valign="middle" style="border-bottom:solid thin" rowspan="1" colspan="1">MeOH</td><td align="center" valign="middle" style="border-bottom:solid thin" rowspan="1" colspan="1">Apocynaceae</td><td align="center" valign="middle" style="border-bottom:solid thin" rowspan="1" colspan="1">Whole plant above ground</td></tr></tbody></table></table-wrap></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
EXPLOR_STEP=$WICRI_ROOT/Sante/explor/ChloroquineV1/Data/Pmc/Curation
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