Fig. 4

YAP rescues PAR2 deficiency-induced apoptosis and colony formation. Different forms of YAP (WT, S127A or S397A) were transfected into stable transfectant HT-29 cells with shRNA against PAR2. a The levels of p127-YAP, p397-YAP, and total-YAP were measured with western blot. b Heatmap showing the 464 rescued genes upon YAP replenishment (shPAR2 + YAP) in shPAR2 HT-29 cells (shPAR2 + vector). Some of the genes related to ROS production (black) and target genes of YAP (CYR61 and GLI2) (red) were listed. c and d HT-29–shPAR2 cells stably expressing YAP (WT), YAP-S127A, YAP-S397A, or vector were cultured in type I collagen and were analyzed for colony formation (5000 cells/well in 12-well plate). c Images of representative field (magnification, 250 ×). d Number of colony was quantified from five consecutive field of each group. e and f ROS was analyzed with flow cytometry in HT-29 stably transfect cells after treatment with H₂O₂ at 1 mM for 2 h. The relative change was obtained by H₂O₂ vs. control group. g and h Apoptosis was analyzed with Annexin V/PI staining assay in HT-29 stably transfect cells after treatment with NO donor DETA NONOate at 0.5 mM for 48 h. The fold change was obtained by NONOate vs. control samples. Data are shown as mean ± SEM. ***p < 0.001 vs. corresponding control