Fig. 3

High AFAP1L1 expression in GC promotes proliferation, invasion in vitro and growth, metastasis in vivo. A The cell proliferative curves were plotted by CCK8 assays for AGS^shAFAP1L1, MKN74^AFAP1L1 and their control cells. B The representative images and the quantification of colony formation assays from AGS^shAFAP1L1, MKN74^AFAP1L1 and their control cells. C Edu assays showed the proliferating cells in AGS^shAFAP1L1, MKN74^AFAP1L1 and their control groups. The percentage of proliferating cells was compared in corresponding bar graphs. Scale bars, 50 μm. D Double fluorescent staining of cytoskeleton (red) and AFAP1L1 (green) for indicated GC cells. Cytoskeleton was stained by rhodamine phalloidin and Nuclei was stained with DAPI (blue). Scale bars, 10 μm. E, F The influence of AFAP1L1 overexpression or knockdown on GC cells migratory and invasive capacity was determined by transwell migration (E) and invasion assays (F). Scale bars, 50 μm. G Subcutaneous tumor model was constructed to test the effect of AFAP1L1 overexpression or knockdown on GC cells growth in vivo. The final tumor volumes were compared in right bar graphs. H In vivo metastatic assays by tail vein injection of indicated GC cells and representative H&E staining images of pulmonary and intrahepatic metastases. FC: fold change. *P < 0.05; **P < 0.01; ***P < 0.001