Fig. 7
AFAP1L1 interacts with VAV2 and activates CDC42 signaling to promote ITGA5 expression. A The protein–protein interaction network of AFAP1L1 was constructed in STRING database (http://string-db.org). Among those interacted proteins, VAV2 attracted our attention. B Co-immunoprecipitation (Co-IP) analysis of the interaction between AFAP1L1 and VAV2 in AGS cells. C VAV2 knockdown and overexpression efficacy in indicated GC cells were determined by qRT-PCR and western blot. D Active CDC42 pull-down assays were performed to detect the level of active CDC42 in AFAP1L1-interfered GC cells with VAV2 further overexpression or knockdown. E The representative IHC images from serial sections of GC samples showed AFAP1L1 and active CDC42 expression in GC tissues with high or low AFAP1L1 expression. Scatter diagram of IHC staining scores showed the expression correlation between AFAP1L1 and active CDC42 in GC. Pearson correlation analysis was utilized to calculate correlation coefficient. F qRT-PCR analysis of ITGA5 and western blot analysis of ITGA5, FAK and ERK in AFAP1L1-interfered GC cells with VAV2 further overexpression or knockdown. ML141 (20 μm) was used to inhibit activation of CDC42 in MKN74AFAP1L1 cells. G Transwell invasion assays were performed to determine the role of VAV2 in mediating the influence of AFAP1L1 on GC cells invasive ability. ML141 (20 μm) was used to inhibit activation of CDC42 in MKN74AFAP1L1 cells. Scale bars, 50 μm. n.s, no significance; *P < 0.05; **P < 0.01; ***P < 0.001