Fig. 2: ERK5 is required for the steady-state YAP transcriptional activity.

A Luciferase assay. MEF2-luc or 8xGTIIC-luc reporters were transiently co-transfected in RLSCs, together with a Renilla expression vector. Luciferase activities were normalized for Renilla luciferase activity and expressed as arbitrary units. Twenty-four hours post-transfection, cells were treated with 10 µM XMD8-92 or its solvent DMSO for 16 h. Statistically significant differences are reported (**p < 0.01; ***p < 0.001). B RT-qPCR analysis of the indicated genes in RLSCs treated with XMD8-92 or DMSO. The values are calculated by the 2(−ΔCt) method, expressed as fold change in gene expression versus the control (DMSO, arbitrary value = 1) and shown as means ± S.E.M. of at least three independent experiments. Statistically significant differences are reported (*p < 0.05; **p < 0.01). C Luciferase assay. MEF2-luc or 8xGTIIC-luc reporters were transiently co-transfected in RLSCs, together with a Renilla expression vector and siERK5 or siGFP. Luciferase activities were normalized for Renilla luciferase activity and expressed as arbitrary units. Statistically significant differences are reported (*p < 0.05). D RT-qPCR analysis of the indicated genes in RLSCs stably transfected with pSUPER or pSUPER-shERK5 vector. The values are calculated by the 2(−ΔCt) method, expressed as fold change in gene expression versus the control (pSUPER, arbitrary value = 1) and shown as means ± S.E.M. of at least three independent experiments. Statistically significant differences are reported (**p < 0.01). E Western blot for the indicated proteins in RLSCs as in (A), (C), and (D). GAPDH has been utilized as loading control. WB images represent one indicative experiment of at least three independent ones.