Fig. 5

YAP/AP-1/ZEB1 complex is involved in ET-1-induced EMT, cellular plasticity, and cell invasion. A E-cadherin, N-cadherin and Vimentin gene expression in OVCAR-3 cells transfected with SCR, si-ZEB1, si-YAP, or si-JUN for 72 h and stimulated with ET-1 and/or treated with macitentan for 24 h as indicated was analyzed by q-RT-PCR and normalized to cyclophilin-A. Values are the means ± SD expressed as fold over control (n = 3; **p < 0.01; #: p value was calculated vs. SCR + ET-1). B Total extracts of OVCAR-3 cells transfected for 72 h and stimulated for 48 h as in A were IB with anti-E-cadherin, anti-N-cadherin or anti-Vimentin Abs. β-actin was used as loading control. C Vasculogenic mimicry assay with OVCAR-3 cells transfected for 48 h and overnight stimulated as in A (Magnification: 20x; scale bar: 100 μm). Right graphs represent the quantification of the number of nodes and the tube length. Columns show the mean ± SD (n = 3; *p < 0.05; **p < 0.01; #: p value was calculated vs. SCR + ET-1). D Transwell chemoinvasion assay with OVCAR-3 cells transfected as in A and overnight allowed to invade in presence of ET-1 and/or macitentan. Images represent the crystal violet-stained invasive cells (Magnification: 20x; scale bar: 100 μm). Right graph represents the number of invading cells. Values are the means ± SD expressed as fold over control (n = 3; *p < 0.05; **p < 0.01; #: p value was calculated vs. SCR + ET-1)