Fig. 6: circMAST1 modulates the proliferation, cell cycle, migration, and invasion of HCC cells via miR-1299.

a Three potential target miRNAs of circMAST1 were predicted by miRANDA, RNAhybrid and regRNA. b Schematic drawing showing the miRNAs that might bind circMAST1. c The possible binding sites of the 5’UTR of G3BP2 with miR-1299 were predicted by RegRNA2.0. Graph of predicted RNA secondary structure. The yellow region indicates the RNA-fold predicted structure of the motif. RNAfold reliability information of pair probabilities. Minimum free energy = −34.02. d Relative Expression of miR-1299 in HCC and normal tissue pairs were measured by qRT-PCR (***P < 0.001; n = 10). e Pearson’s correlation analyses showing the correlation of circMAST1 and miR-1299 expression (n = 10). f Schematic of circMAST1 wild type(wt) and mutant(mut) luciferase reporter vectors. luciferase reporter assay in HEK293T cells co-transfected with miRNA mimics, circMAST1 wild type (circMAST1-wt) and mutant (mut) luciferase reporter vectors. g The expression of miR-1299 were analyzed by using qRT-PCR in cells transfected with siRNA-circMAST1-1, siRNA-circMAST1-3 or siRNA-NC (**P < 0.01; n = 6). h, i The viability of HCLLM3 and hepG2 cells were measured after co-transfected siRNA-circMAST1 and miR-1299 inhibitor in by using WST-1 assays (*P < 0.05; **P < 0.01; n = 4). j Cell proliferation ability of HCLLM3 and hepG2 cells co-transfected with siRNA-circMAST1-1 and miR-1299 inhibitors were evaluated by colony formation assay (*P < 0.05; **P < 0.01; n = 4). k, l The expression of PCNA, cyclin A, cyclin E and CDK1, CDK2 in HCLLM3 and hepG2 cells by Western-blot. Bar graph showed the results of the expression of PCNA, cyclin A, cyclin D and CDK1, CDK2 in HCLLM3 and hepG2 cells (*P < 0.05; **P < 0.01; ***P < 0.001; n = 4). m, n Cell migration or invasion assays were performed in HCLLM3 and hepG2 cells co-transfected with siRNA-circMAST1-1 and miR-1299 inhibitors by using transwell chamber with or without matrigel respectively (**P < 0.01; n = 4).