Fig. 3: ERK5 is required for endogenous and exogenous YAP transcriptional activity in HuH7.

A RT-qPCR analysis of the indicated genes in HuH7 treated with 10 µM BIX02189 or DMSO. Data are expressed as relative gene expression and shown as mean ± S.E.M. of three independent experiments. Statistically significant differences are reported (*p < 0.05). B Western blot for the indicated proteins in HuH7 as in (A). GAPDH has been utilized as loading control. C RT-qPCR analysis of the indicated genes in HuH7 transfected with pSUPER or pSUPER-shERK5 vector. Data are expressed as in (A). Statistically significant differences are reported (*p < 0.05; **p < 0.01; ***p < 0.001). D Western blot for the indicated proteins in HuH7 as in (C). GAPDH has been utilized as loading control. E RT-qPCR analysis of the indicated genes in HuH7 cells, grown as spheroids for 24 h and treated with 10 µM BIX02189 or DMSO. Data are expressed as relative gene expression and shown as mean ± S.E.M. of three independent 3D cell cultures. Statistically significant differences are reported (unpaired, one-tailed Student’s t test; *p < 0.05; ns= not significant). Representative images of spheroids in the two different cell conditions are shown. F RT-qPCR analysis of the indicated YAP target genes in wild-type YAP-overexpressing HuH7 cells (YAP-WT) and in control cells (CTR), treated with 10 µM BIX02189 or DMSO. Data are expressed as relative gene expression and shown as mean ± S.E.M. of three independent experiments. Statistically significant differences are reported (**p < 0.01; ***p < 0.001).