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. 2019 Aug 30;10:942. doi: 10.3389/fphar.2019.00942

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Copyright © 2019 Bisceglia, Battistelli, Noce, Montaldo, Zammataro, Strippoli, Tripodi, Amicone and Marchetti

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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TGFβ overrides HNF1α constitutive expression. (A) Analysis of constitutively expressed HNF1α activity following TGFβ treatment. RT-qPCR analysis for the indicated genes in hepatocytes, transiently transfected with pLCPX-HNF1α^Myc (HNF1) or the empty vector (CTR), treated with 4 ng/ml TGFβ for 24 h or left untreated (NT). qPCR data, obtained in triplicate and normalized to the housekeeping gene RPL34, are expressed as relative gene expression. The mean ± SEM of three independent experiments is shown. Statistically significant differences are reported (*p < 0.05; ns, not significant). (B) Immunofluorescence analysis of hepatocytes transfected and treated as in (A). Cells were stained with anti-MycTag antibody (red) and 4’,6-diamidino-2-phenylindole (nuclei, blue). Magnification 10×. (C) Analysis of EMT-related gene expression by RT-qPCR in hepatocytes, transiently transfected with pLCPX-HNF1αMyc (HNF1) or the empty vector (CTR), treated with TGFβ for 24 h or left untreated (NT). qPCR data, obtained in triplicate and normalized to the housekeeping gene RPL34, are expressed as relative gene expression. The mean ± SEM of three independent experiments is shown. Statistically significant differences are reported (*p < 0.05; ns, not significant). (D) Immunofluorescence analysis of hepatocytes transfected and treated as in (C). Cells were stained with anti-E-cadherin (green) or anti-HNF1α antibody (red). Magnification 20×.