Fig. 3

Activation of PAR2 promotes YAP stability. HEK293 cells were serum-starved for 12 h and then stimulated with (a) trypsin (1.5 nM or 2.5 nM) or (b) PAR2-activating peptide (AP) (100 μM) for different times. The level of total YAP, p127-YAP, and p397-YAP were measured with western blot. c HEK293 cells were serum-starved for 12 h and then stimulated with PAR2-AP (100 μM) or trypsin (2.5 nM) for 1 h. Expression of YAP mRNA was measured with real-time PCR. d Western blot was used to measure YAP protein expression in the nuclear and cytoplasmic fractions from HEK293 cell after PAR2 activation for different time. PARP (nuclear marker) and Hsp90 (cytoplasmic marker) were detected. e mRNA expression of YAP target genes were measured with real-time PCR. f Protein levels of YAP and PAR2 were examined with western blot in stable transfectant HT-29 cells with or without shRNA against PAR2 (sh-PAR2). g Expression of F2RL1 (PAR2), YAP, and (h) its target genes were examined with real-time PCR in stable transfectant HT-29 cells. i p127-YAP, p397-YAP, and total-YAP were measured with western blot in stable transfectant HT-29 cells. Note: To have equal amount of YAP protein, the lysates of HT-29-vector and HT-29-shPAR2 are loaded at the ratio of 1:2.5. Data were shown as mean ± SEM. N.S. no significance; *p < 0.05; **p < 0.01; ***p < 0.001 vs. corresponding control