Fig. 5

GRK3 positively regulated YAP1 targets and GRK3 inhibitor reduced CSC attributes and aggressive phenotypes of GAC cells. A SOX9, Birc5, Cyr61 and CTGF were detected by qPCR in KATO III cells with or without GRK3 OE; **P < 0.001. B SOX9, Birc5, Cyr61 and CTGF were detected by qPCR in MKN45 cells with or without GRK3 OE; ***P < 0.0001.C SOX9 expression was detected by Western blot (upper panel) or co-IF in both KATO III and MKN45 cells with or without GRK3 cDNA overexpression (Bottom panel). Scale bar: 50µm. D The level of YAP1 and its targets SOX9, Cyr61 were detected using qPCR in GA0518 and GA0804 patient derived cells treated with or without LD2 at dosage indicated. ****P < 0.0001. E Level of SOX9 was determined by Western blot (upper panel) and co-IF (bottom panel) in GA0518 cells and treated with LD2 at indicated dosage. Scale bar: 50µm. F Tumor sphere formation was detected in MKN45 and KATO III cells with or without overexpression of GRK3 and then treated with LD2 at indicated dosage. *P < 0.01, **P < 0.001, ***P < 0.0001. G Representative image (left) and quantification (right) of ALDH1A1⁺ population was labeled using the ALDEFLUOR kit in KATO III cells with GRK3 OE compared to the control (n = 3 biological replicates). P = 0.000424. H Representative image (left) and quantification (right) of ALDH1A1⁺ population was labeled using the ALDEFLUOR kit in GA0518 cells with LD2 treatment at 0.5µM for 48 h (n = 3 biological replicates). P = 0.000741