Fig. 7
From: Regulation of the Hippo/YAP axis by CXCR7 in the tumorigenesis of gastric cancer
CXCR7 activates YAP through the Gαq/11-ROCK-LATS axis in gastric cancer. A, B CXCR7 activation induced YAP dephosphorylation through Gaq/11. MGC803 and Hs746T cells were transiently transfected with control, Gaq/11, or Gas siRNAs. MGC803 and Hs746T cells were treated with TC. YAP, phosphorylated YAP, Gaq/11 and Gas were determined by immunoblotting. C CXCR7 activation induces YAP nuclear localization through Gaq/11. MGC803 cells were transiently transfected with control, Gaq/11 or Gas siRNAs. MGC803 cells were treated with TC. Endogenous YAP (green) and nuclei (blue) were stained with specific antibodies and DAPI, respectively; scale bar, 20 mm. Quantifications of YAP subcellular localization from at least 100 randomly selected cells. C, cytoplasm; N, nucleus. D Nucleoplasm separation experiments by immunoblotting confirmed that Gαq/11 silencing could block the YAP nuclear accumulation caused by CXCR7 activation in MGC 803 cells. E CXCR7 activation via TC decreases LATS1 activity. MGC803 and Hs746T cells were stimulated with TC. LATS1 was immunoprecipitated. Phosphorylation of YAP by LATS1 was determined by a phospho-YAP antibody. F Ectopic expression of LATS1 blocks YAP dephosphorylation induced by CXCR activation. MGC803 and Hs746T cells were transiently transfected with control, LATS1 wild type (WT), or kinase dead mutant (K/R). MGC803 and Hs746T cells were treated with TC for 1 h. Phosphorylation and protein levels of YAP were determined by immunoblotting. G Rho GTPase is involved in YAP dephosphorylation induced by CXCR activation. MGC803 and Hs746T cells were transiently transfected with control, Myc-Rho-L63, or C3. MGC803 and Hs746T cells were treated with TC. Total YAP and phosphorylated YAP protein levels were determined by immunoblotting. H ROCK is required for CXCR7-induced YAP activation. Serum-starved MGC803 and Hs746T cells were pretreated with GSK429286 (1 mmol/L) or Y27632 (1 mmol/L) for 4 h, followed by treatment with TC. Total YAP and phosphorylated YAP protein levels were determined by immunoblotting