Fig. 6

GNB4 activates downstream Hippo-YAP1 pathway molecules in GC cells. A GSEA was based on GNB4 expression in patients with GC (data obtained from the TCGA database) using CAMOIP online tool. B, C qRT-PCR analysis of GNB4, YAP1, TAZ, CYR61, and CTGF expression in MKN45 (B) and AGS (C) cells with or without GNB4 expression. D Expression of E-cadherin (E-cad), N-cadherin (N-cad), and the indicated Hippo pathway molecules (YAP1, TEAD1/2/3/4, CYR61, and CTGF) in AGS and MKN45 cells without and with GNB4 overexpression (measured by using western blotting). E Expression of GNB4 and YAP1 in MKN45 and AGS cells treated with increasing concentrations of verteporfin (VP) for 12 h (using western blot assay). F, G AGS cells were transfected with empty vector and GNB4-overexpression lentivirus. Nuclear and cytoplasmic protein extraction and western blotting of GNB4 and YAP1 were performed (F). Representative images showing immunofluorescence staining (scale bar: 20 μm) for YAP1 (green) (G). H Immunoprecipitation assay of AGS cells with and without GNB4 overexpression using an anti-YAP1 antibody. Immunoglobulin G (IgG) was used as a negative control. Western blotting was performed to determine YAP1, TEAD1/2/3/4, and GNB4 expression