Fig. 4: ERK5 constitutive activation promotes YAP transcriptional activity. | Cell Death & Disease

Fig. 4: ERK5 constitutive activation promotes YAP transcriptional activity.

From: Extracellular signal-Regulated Kinase 5 (ERK5) is required for the Yes-associated protein (YAP) co-transcriptional activity

Fig. 4

A Luciferase assay. MEF2-luc or 8xGTIIC-luc reporters were transiently co-transfected in HepE14 cells, together with a Renilla expression vector, an ERK5 and a constitutive active MEK5 expressing vector (ERK5/caMEK5) or the empty vector (pcDNA3). Luciferase activities were normalized for Renilla luciferase activity and expressed as arbitrary units. Statistically significant differences are reported (*p < 0.05; **p < 0.01). B Western blot for the indicated proteins in ERK5/caMEK5-overexpressing HepE14. GAPDH has been utilized as loading control. WB images represent one indicative experiment of three independent ones. C RT-qPCR analysis of the indicated YAP target genes in ERK5/caMEK5-overexpressing RLSCs. The values are calculated by the 2(−ΔCt) method, expressed as fold change in gene expression versus the control (empty vector, 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). D RT-qPCR analysis of the indicated YAP target genes in ERK5/caMEK5-overexpressing and in control HuH7 cells, treated with 10 µM of YAP-TEAD inhibitor Verteporfin (VP) or with DMSO. The values are calculated by the 2(−ΔCt) method and shown as means ± S.E.M. of three independent experiments. Statistically significant differences are reported (*p < 0.05; **p < 0.01). E RT-qPCR analysis of the indicated YAP target genes in YAP-silenced ERK5/caMEK5-overexpressing HuH7 cells (siYAP), compared with cells transfected with control siRNAs (siCTR). The values are calculated by the 2(−ΔCt) method and shown as means ± S.E.M. of three independent experiments. Statistically significant differences are reported (*p < 0.05; **p < 0.01). F Luciferase assay. 8xGTIIC-luc reporter was transiently co-transfected in HuH7 cells, together with a Renilla expression vector, an ERK5 and a constitutive active MEK5 expressing vector (ERK5/caMEK5) or the empty vector (pcDNA3), in the presence of siYAP or siCTR. Luciferase activities were normalized for Renilla luciferase activity and expressed as arbitrary units. Statistically significant differences are reported (*p < 0.05; **p < 0.01).

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