Induction of highly functional hepatocytes from human umbilical cord mesenchymal stem cells by HNF4α transduction
To investigate the differentiation potential of human umbilical mesenchymal stem cells (HuMSCs) and the key factors that facilitate hepatic differentiation. HuMSCs were induced to become hepatocyte-like cells according to a previously published protocol. The differentiation status of the hepatocyte-...
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| Veröffentlicht in: | PloS one 2014-08, Vol.9 (8), p.e104133-e104133 |
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| creator | Hang, Hualian Yu, Yabin Wu, Ning Huang, Qingfeng Xia, Qiang Bian, Jianmin |
| description | To investigate the differentiation potential of human umbilical mesenchymal stem cells (HuMSCs) and the key factors that facilitate hepatic differentiation.
HuMSCs were induced to become hepatocyte-like cells according to a previously published protocol. The differentiation status of the hepatocyte-like cells was examined by observing the morphological changes under an inverted microscope and by immunofluorescence analysis. Hepatocyte nuclear factor 4 alpha (HNF4α) overexpression was achieved by plasmid transfection of the hepatocyte-like cells. The expression of proteins and genes of interest was then examined by Western blotting and reverse transcription-polymerase chain reaction (RT-PCR) or real-time RT-PCR methods.
Our results demonstrated that HuMSCs can easily be induced into hepatocyte-like cells using a published differentiation protocol. The overexpression of HNF4α in the induced HuMSCs significantly enhanced the expression levels of hepatic-specific proteins and genes. HNF4α overexpression may be associated with liver-enriched transcription factor networks and the Wnt/β-Catenin pathway.
The overexpression of HNF4α improves the hepatic differentiation of HuMSCs and is a simple way to improve cellular sources for clinical applications. |
| doi_str_mv | 10.1371/journal.pone.0104133 |
| format | Article |
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HuMSCs were induced to become hepatocyte-like cells according to a previously published protocol. The differentiation status of the hepatocyte-like cells was examined by observing the morphological changes under an inverted microscope and by immunofluorescence analysis. Hepatocyte nuclear factor 4 alpha (HNF4α) overexpression was achieved by plasmid transfection of the hepatocyte-like cells. The expression of proteins and genes of interest was then examined by Western blotting and reverse transcription-polymerase chain reaction (RT-PCR) or real-time RT-PCR methods.
Our results demonstrated that HuMSCs can easily be induced into hepatocyte-like cells using a published differentiation protocol. The overexpression of HNF4α in the induced HuMSCs significantly enhanced the expression levels of hepatic-specific proteins and genes. HNF4α overexpression may be associated with liver-enriched transcription factor networks and the Wnt/β-Catenin pathway.
The overexpression of HNF4α improves the hepatic differentiation of HuMSCs and is a simple way to improve cellular sources for clinical applications.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0104133</identifier><identifier>PMID: 25137413</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Activins - pharmacology ; Adipocytes - cytology ; Adipocytes - drug effects ; Adipocytes - metabolism ; beta Catenin - genetics ; beta Catenin - metabolism ; Biology and Life Sciences ; Cell Differentiation ; Cells, Cultured ; Chondrocytes - cytology ; Chondrocytes - drug effects ; Chondrocytes - metabolism ; Differentiation ; Embryos ; Epidermal Growth Factor - pharmacology ; Fetal Blood - cytology ; Fetal Blood - drug effects ; Fetal Blood - metabolism ; Gene Expression ; Genes ; Hepatocyte nuclear factor 4 ; Hepatocyte Nuclear Factor 4 - genetics ; Hepatocyte Nuclear Factor 4 - metabolism ; Hepatocytes ; Hepatocytes - cytology ; Hepatocytes - drug effects ; Hepatocytes - metabolism ; Humans ; Immunofluorescence ; Insulin - pharmacology ; Liver ; Medicine and Health Sciences ; Mesenchymal stem cells ; Mesenchymal Stromal Cells - cytology ; Mesenchymal Stromal Cells - drug effects ; Mesenchymal Stromal Cells - metabolism ; Mesenchyme ; Niacinamide - pharmacology ; Plasmids ; Polymerase chain reaction ; Proteins ; Reverse transcription ; Rodents ; Selenium - pharmacology ; Signal Transduction ; Skin & tissue grafts ; Stem cells ; Therapeutic applications ; Transfection ; Transferrin - pharmacology ; Umbilical cord ; Western blotting ; Wnt protein ; Wnt Proteins - genetics ; Wnt Proteins - metabolism ; β-Catenin</subject><ispartof>PloS one, 2014-08, Vol.9 (8), p.e104133-e104133</ispartof><rights>2014 Hang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2014 Hang et al 2014 Hang et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c526t-859cb93d4cc5ddfef3e63b2c631dc3b72ccad0423ba4e478449f33c124692e013</citedby><cites>FETCH-LOGICAL-c526t-859cb93d4cc5ddfef3e63b2c631dc3b72ccad0423ba4e478449f33c124692e013</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4138090/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4138090/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,864,885,2102,2928,23866,27924,27925,53791,53793,79600,79601</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25137413$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Almeida-Porada, Graca</contributor><creatorcontrib>Hang, Hualian</creatorcontrib><creatorcontrib>Yu, Yabin</creatorcontrib><creatorcontrib>Wu, Ning</creatorcontrib><creatorcontrib>Huang, Qingfeng</creatorcontrib><creatorcontrib>Xia, Qiang</creatorcontrib><creatorcontrib>Bian, Jianmin</creatorcontrib><title>Induction of highly functional hepatocytes from human umbilical cord mesenchymal stem cells by HNF4α transduction</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>To investigate the differentiation potential of human umbilical mesenchymal stem cells (HuMSCs) and the key factors that facilitate hepatic differentiation.
HuMSCs were induced to become hepatocyte-like cells according to a previously published protocol. The differentiation status of the hepatocyte-like cells was examined by observing the morphological changes under an inverted microscope and by immunofluorescence analysis. Hepatocyte nuclear factor 4 alpha (HNF4α) overexpression was achieved by plasmid transfection of the hepatocyte-like cells. The expression of proteins and genes of interest was then examined by Western blotting and reverse transcription-polymerase chain reaction (RT-PCR) or real-time RT-PCR methods.
Our results demonstrated that HuMSCs can easily be induced into hepatocyte-like cells using a published differentiation protocol. The overexpression of HNF4α in the induced HuMSCs significantly enhanced the expression levels of hepatic-specific proteins and genes. HNF4α overexpression may be associated with liver-enriched transcription factor networks and the Wnt/β-Catenin pathway.
The overexpression of HNF4α improves the hepatic differentiation of HuMSCs and is a simple way to improve cellular sources for clinical applications.</description><subject>Activins - pharmacology</subject><subject>Adipocytes - cytology</subject><subject>Adipocytes - drug effects</subject><subject>Adipocytes - metabolism</subject><subject>beta Catenin - genetics</subject><subject>beta Catenin - metabolism</subject><subject>Biology and Life Sciences</subject><subject>Cell Differentiation</subject><subject>Cells, Cultured</subject><subject>Chondrocytes - cytology</subject><subject>Chondrocytes - drug effects</subject><subject>Chondrocytes - metabolism</subject><subject>Differentiation</subject><subject>Embryos</subject><subject>Epidermal Growth Factor - pharmacology</subject><subject>Fetal Blood - cytology</subject><subject>Fetal Blood - drug effects</subject><subject>Fetal Blood - metabolism</subject><subject>Gene Expression</subject><subject>Genes</subject><subject>Hepatocyte nuclear factor 4</subject><subject>Hepatocyte Nuclear Factor 4 - genetics</subject><subject>Hepatocyte Nuclear Factor 4 - metabolism</subject><subject>Hepatocytes</subject><subject>Hepatocytes - cytology</subject><subject>Hepatocytes - drug effects</subject><subject>Hepatocytes - metabolism</subject><subject>Humans</subject><subject>Immunofluorescence</subject><subject>Insulin - pharmacology</subject><subject>Liver</subject><subject>Medicine and Health Sciences</subject><subject>Mesenchymal stem cells</subject><subject>Mesenchymal Stromal Cells - cytology</subject><subject>Mesenchymal Stromal Cells - drug effects</subject><subject>Mesenchymal Stromal Cells - metabolism</subject><subject>Mesenchyme</subject><subject>Niacinamide - pharmacology</subject><subject>Plasmids</subject><subject>Polymerase chain reaction</subject><subject>Proteins</subject><subject>Reverse transcription</subject><subject>Rodents</subject><subject>Selenium - pharmacology</subject><subject>Signal Transduction</subject><subject>Skin & tissue grafts</subject><subject>Stem cells</subject><subject>Therapeutic applications</subject><subject>Transfection</subject><subject>Transferrin - pharmacology</subject><subject>Umbilical cord</subject><subject>Western blotting</subject><subject>Wnt protein</subject><subject>Wnt Proteins - genetics</subject><subject>Wnt Proteins - metabolism</subject><subject>β-Catenin</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><sourceid>DOA</sourceid><recordid>eNptUstu1DAUjRCIlsIfILDEppsZbF_H42yQUNXSkSrYwNpybGeSkWMPdoKUz-JH-CacmbRqEStb955z7usUxVuC1wQ25OM-jNErtz4Eb9eYYEYAnhXnpAK64hTD80f_s-JVSnuMSxCcvyzOaJklMuG8iFtvRj10waPQoLbbtW5CzeiPIeVQaw9qCHoabEJNDD1qx155NPZ15zqdATpEg3qbrNft1OdAGmyPtHUuoXpCt19v2J_faIjKp6XQ6-JFo1yyb5b3ovhxc_396nZ19-3L9urz3UqXlA8rUVa6rsAwrUtjGtuA5VBTzYEYDfWGaq0MZhRqxSzbCMaqBkATynhFLSZwUbw_6R5cSHJZV5KkLFlJKAeWEdsTwgS1l4fY9SpOMqhOHgMh7qSKQ6edlRQ4s9jUfCMqpsEK0fC5PwGiNKKaq31aqo11b422Ps_snog-zfiulbvwS-YzCFzhLHC5CMTwc7RpkH2X5j0qb8N47LvMtxR4k6Ef_oH-fzp2QukYUoq2eWiGYDlb6J4lZwvJxUKZ9u7xIA-ke8_AX_rDxvQ</recordid><startdate>20140819</startdate><enddate>20140819</enddate><creator>Hang, Hualian</creator><creator>Yu, Yabin</creator><creator>Wu, Ning</creator><creator>Huang, Qingfeng</creator><creator>Xia, Qiang</creator><creator>Bian, Jianmin</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20140819</creationdate><title>Induction of highly functional hepatocytes from human umbilical cord mesenchymal stem cells by HNF4α transduction</title><author>Hang, Hualian ; Yu, Yabin ; Wu, Ning ; Huang, Qingfeng ; Xia, Qiang ; Bian, Jianmin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c526t-859cb93d4cc5ddfef3e63b2c631dc3b72ccad0423ba4e478449f33c124692e013</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Activins - pharmacology</topic><topic>Adipocytes - cytology</topic><topic>Adipocytes - drug effects</topic><topic>Adipocytes - metabolism</topic><topic>beta Catenin - genetics</topic><topic>beta Catenin - metabolism</topic><topic>Biology and Life Sciences</topic><topic>Cell Differentiation</topic><topic>Cells, Cultured</topic><topic>Chondrocytes - cytology</topic><topic>Chondrocytes - drug effects</topic><topic>Chondrocytes - metabolism</topic><topic>Differentiation</topic><topic>Embryos</topic><topic>Epidermal Growth Factor - pharmacology</topic><topic>Fetal Blood - cytology</topic><topic>Fetal Blood - drug effects</topic><topic>Fetal Blood - metabolism</topic><topic>Gene Expression</topic><topic>Genes</topic><topic>Hepatocyte nuclear factor 4</topic><topic>Hepatocyte Nuclear Factor 4 - genetics</topic><topic>Hepatocyte Nuclear Factor 4 - metabolism</topic><topic>Hepatocytes</topic><topic>Hepatocytes - cytology</topic><topic>Hepatocytes - drug effects</topic><topic>Hepatocytes - metabolism</topic><topic>Humans</topic><topic>Immunofluorescence</topic><topic>Insulin - pharmacology</topic><topic>Liver</topic><topic>Medicine and Health Sciences</topic><topic>Mesenchymal stem cells</topic><topic>Mesenchymal Stromal Cells - cytology</topic><topic>Mesenchymal Stromal Cells - drug effects</topic><topic>Mesenchymal Stromal Cells - metabolism</topic><topic>Mesenchyme</topic><topic>Niacinamide - pharmacology</topic><topic>Plasmids</topic><topic>Polymerase chain reaction</topic><topic>Proteins</topic><topic>Reverse transcription</topic><topic>Rodents</topic><topic>Selenium - pharmacology</topic><topic>Signal Transduction</topic><topic>Skin & tissue grafts</topic><topic>Stem cells</topic><topic>Therapeutic applications</topic><topic>Transfection</topic><topic>Transferrin - pharmacology</topic><topic>Umbilical cord</topic><topic>Western blotting</topic><topic>Wnt protein</topic><topic>Wnt Proteins - genetics</topic><topic>Wnt Proteins - metabolism</topic><topic>β-Catenin</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hang, Hualian</creatorcontrib><creatorcontrib>Yu, Yabin</creatorcontrib><creatorcontrib>Wu, Ning</creatorcontrib><creatorcontrib>Huang, Qingfeng</creatorcontrib><creatorcontrib>Xia, Qiang</creatorcontrib><creatorcontrib>Bian, Jianmin</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Meteorological & Geoastrophysical Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Collection</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Meteorological & Geoastrophysical Abstracts - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hang, Hualian</au><au>Yu, Yabin</au><au>Wu, Ning</au><au>Huang, Qingfeng</au><au>Xia, Qiang</au><au>Bian, Jianmin</au><au>Almeida-Porada, Graca</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Induction of highly functional hepatocytes from human umbilical cord mesenchymal stem cells by HNF4α transduction</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2014-08-19</date><risdate>2014</risdate><volume>9</volume><issue>8</issue><spage>e104133</spage><epage>e104133</epage><pages>e104133-e104133</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>To investigate the differentiation potential of human umbilical mesenchymal stem cells (HuMSCs) and the key factors that facilitate hepatic differentiation.
HuMSCs were induced to become hepatocyte-like cells according to a previously published protocol. The differentiation status of the hepatocyte-like cells was examined by observing the morphological changes under an inverted microscope and by immunofluorescence analysis. Hepatocyte nuclear factor 4 alpha (HNF4α) overexpression was achieved by plasmid transfection of the hepatocyte-like cells. The expression of proteins and genes of interest was then examined by Western blotting and reverse transcription-polymerase chain reaction (RT-PCR) or real-time RT-PCR methods.
Our results demonstrated that HuMSCs can easily be induced into hepatocyte-like cells using a published differentiation protocol. The overexpression of HNF4α in the induced HuMSCs significantly enhanced the expression levels of hepatic-specific proteins and genes. HNF4α overexpression may be associated with liver-enriched transcription factor networks and the Wnt/β-Catenin pathway.
The overexpression of HNF4α improves the hepatic differentiation of HuMSCs and is a simple way to improve cellular sources for clinical applications.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>25137413</pmid><doi>10.1371/journal.pone.0104133</doi><oa>free_for_read</oa></addata></record> |
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| subjects | Activins - pharmacology Adipocytes - cytology Adipocytes - drug effects Adipocytes - metabolism beta Catenin - genetics beta Catenin - metabolism Biology and Life Sciences Cell Differentiation Cells, Cultured Chondrocytes - cytology Chondrocytes - drug effects Chondrocytes - metabolism Differentiation Embryos Epidermal Growth Factor - pharmacology Fetal Blood - cytology Fetal Blood - drug effects Fetal Blood - metabolism Gene Expression Genes Hepatocyte nuclear factor 4 Hepatocyte Nuclear Factor 4 - genetics Hepatocyte Nuclear Factor 4 - metabolism Hepatocytes Hepatocytes - cytology Hepatocytes - drug effects Hepatocytes - metabolism Humans Immunofluorescence Insulin - pharmacology Liver Medicine and Health Sciences Mesenchymal stem cells Mesenchymal Stromal Cells - cytology Mesenchymal Stromal Cells - drug effects Mesenchymal Stromal Cells - metabolism Mesenchyme Niacinamide - pharmacology Plasmids Polymerase chain reaction Proteins Reverse transcription Rodents Selenium - pharmacology Signal Transduction Skin & tissue grafts Stem cells Therapeutic applications Transfection Transferrin - pharmacology Umbilical cord Western blotting Wnt protein Wnt Proteins - genetics Wnt Proteins - metabolism β-Catenin |
| title | Induction of highly functional hepatocytes from human umbilical cord mesenchymal stem cells by HNF4α transduction |
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