Fig. 2
ET-1/ETAR axis promotes the engagement of the ZEB1/YAP/AP-1 transcriptional nuclear complex. A Nuclear extracts of PMOV10 cells stimulated for 6 h with ET-1 and/or with macitentan as indicated were IP for endogenous YAP using anti-YAP or anti-IgG as control Ab and IB using Abs recognizing the ZEB1, YAP, TEAD4, or c-JUN (JUN) proteins. PCNA was used as loading control. B-C Nuclear extracts of PMOV10 cells transfected with siRNA control (SCR) and si-ZEB1 (B) or si-YAP (C) for 72 h and stimulated for 6 h with ET-1 as indicated were IP using anti-IgG Ab control, anti-YAP Ab for endogenous YAP (B), and/or anti-ZEB1 Ab for endogenous ZEB1 (C), and IB as in A. D Representative images of PLA detection of protein complex containing ZEB1 and JUN (red signals) in PMOV10 cells stimulated or not with ET-1 for 6 h. DAPI staining (blue) highlights the nucleus (Magnification: 63x; scale bar: 10 μm). Right graph represents the quantification of the protein complex. Bars are means ± SD (n = 3; **p < 0.01)