Fig. 3
CCL3 promotes tumor migration and invasion by regulating m6A modification through VIRMA in ICC. a The m6A dot blot assay of global m6A abundance in mRNA of RBE cells co-cultured with THLE3. b The m6A dot blot assay of global m6A abundance in mRNA of RBE cells stimulated with CCL3 (100 ng/ml) for 6 h. c Left: qRT-PCR analysis of m6A-related genes mRNA expression in RBE cells stimulated with CCL3. The relative quantification was calculated using the 2−ΔΔCt method and normalized based on GAPDH. Right: western blot analysis of VIRMA protein expression in RBE cells stimulated with CCL3 (100 ng/ml) for 6 h. d qRT-PCR and western blot analysis of VIRMA expression in RBE cells stimulated with CCL3 (100 ng/ml) for different times (0, 2, 6, 12, and 24). e qRT-PCR and western blot analysis of VIRMA expression in RBE cells stimulated with different concentrations of CCL3 (0, 20, 50, 100, and 200 ng/ml) for 6 h. f After treating with CCR1, CCR4, or CCR5 receptor antagonists, qRT-PCR and western blot analysis of VIRMA expression in RBE cells stimulated with CCL3 (100 ng/ml) for 6 h. g Pro-metastatic effects of the CCL3/VIRMA pathway were confirmed by transwell assays of RBE cells and HuCCT1 cells with different treatments. Scale bars, 100 μm. *P < 0.05 **P < 0.01, ***P < 0.001