ASAP3 disrupts ASAP1-ARHGAP12 to inhibit RhoA and yes-associated protein/transcriptional coactivator with PDZ-binding motif, suppressing gastric cancer progression

Figure 8 Validation of the related mechanisms using real-time quantitative polymerase chain reaction, western blot, co-immunoprecipitation and pull down. A: The messenger RNA expression of CCN1, CCN2 and AMOTL2 in gastric cancer (GC) cells with different treatments measured using real-time quantitative polymerase chain reaction; B: The protein expression of phosphorylated (p)-MST1/MST2, p-LATS1 + LAST2, p-yes-associated protein, p-transcriptional coactivator with PDZ-binding motif, ASAP1 and ARHGAP12 in GC cells with different treatments detected by western blot; C: The relationship between ASAP3 and ASAP1-ARHGAP12 interaction determined by co-immunoprecipitation; D: The correlation between ASAP3 and RhoA activity examined by RhoA activation pull down. ^cP < 0.05 vs short hairpin-negative control-HGC-27 cell. ^dP < 0.05 vs over-expression-negative control-AGS cell. mRNA: Messenger RNA; oe: Over-expression; sh: Short hairpin; NC: Negative control; YAP: Yes-associated protein; TAZ: Transcriptional coactivator with PDZ-binding motif; p-YAP: Phosphorylated yes-associated protein; p-TAZ: Phosphorylated transcriptional coactivator with PDZ-binding motif; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; IP: Immunoprecipitation; WB: Western blot.