Fig. 6: Adhesion- and TGFβ- mediated activation of YAP requires ERK5.

A RT-qPCR analysis of the indicated genes in HuH7. Cells were trypsinized, maintained in suspension for 10’ and collected (Susp) or plated for 3 h on collagen-coated plates (Adh) in the presence or absence of ERK5 inhibitor BIX02189. Data are expressed as relative expression and shown as means ± S.E.M. of at least three independent experiments. Statistically significant differences are reported (*p < 0.05; **p < 0.01). B RT-qPCR analysis of the indicated genes in suspended and adherent YAP-silenced HuH7 (siYAP), compared with cells transfected with control siRNAs (siCTR). Data are expressed as relative expression and shown as means ± S.E.M. of at least three independent experiments. Statistically significant differences are reported (*p < 0.05; ns =not significant). C Luciferase assay. MEF2-luc or 8xGTIIC-luc reporters were transiently co-transfected in HepE14, together with a Renilla luciferase expression vector. Twenty-four hours post-transfection, cells were treated with TGFβ1 or left untreated for 24 h, in the presence of 10 µM XMD8-92 or DMSO. Luciferase activities were normalized for Renilla luciferase activity and expressed as arbitrary units. Statistically significant differences are reported (*p < 0.05; **p < 0.01; ***p < 0.001; ns = not significant). D RT-qPCR analysis of Ctgf and Cyr61 gene expression in untreated or TGFβ1-treated HepE14 cells. The values are calculated by the 2(−ΔCt) method and shown as means ± S.E.M. of at least three independent experiments. Statistically significant differences are reported (**p < 0.01; ***p < 0.001; ns = not significant).