Fig. 4.

YAP1 overexpression increases anoikis resistance and metastasis. a Analysis of YAP1 protein expression in ovarian and colon cancer cell lines. GAPDH was used as a loading control (n = 2). b Validation of overexpression of YAP1 with wild-type (WT) YAP1, constitutively active (S127A) YAP1 and TEAD-defective mutant (S127A/S94A) YAP1 in OVCA432 human ovarian cancer cells. GAPDH was used as a loading control (n = 3). c Cell viability analysis (upper panel) and representative crystal violet staining (lower panel) of OVCA432 cells transduced with control or YAP1 overexpressing constructs 72 h after seeding into 96-well plates (n = 6, one-way ANOVA followed by a Tukey’s multiple comparison post hoc test). d Analysis of GFP-positive, viable (SYTOX Red negative) cells after 72 h of low attachment in OVCA432 cells transduced with control or YAP1 overexpressing constructs (n = 6, one-way ANOVA followed by a Tukey’s multiple comparison post hoc test). e Anchorage-independent growth assay in soft agar using OVCA432 transduced with control or YAP1 overexpressing constructs and quantification of colonies after seeding 15,000 cells and incubation for 14 days (n = 3, one-way ANOVA followed by a Tukey’s multiple comparison post-hoc test). f–h Representative necropsy pictures f, number of metastatic nodules g and weight of ovarian primary tumor h after intraovarian injection of OVCA432 overexpressed with either eGFP-control, YAP^WT, YAP^S127A or YAP^S127A/S94A constructs (n = 10, one-way ANOVA followed by a Tukey’s multiple comparison post-hoc test). Bars and error bars represent mean values and the corresponding SEMs (*p < 0.05, **p < 0.01, ***p < 0.001)