Fig. 2

EVA1A is a direct target gene of miR-103a-3p. A The potential upstream microRNA of EVA1A by microRNA prediction databases. B Ten pairs of samples were randomly selected, RT-qPCR analysis of EVA1A mRNA levels and predicted five microRNA levels. C miR-103a-3p expression was inversely correlated with EVA1A levels according to data from HCC patients in the TCGA cohort (p = 0.001; Pearson correlation). D Scheme of the potential binding sequence of miR-103a-3p and miR-107 within the human EVA1A 3′UTR and construction wild-type (wt) and mutated (mut) luciferase plasmids basing on the EVA1A 3′UTR binding sequence. E Human miR-103a-3p or mimic controls (NC mimics) were co-transfected wt or mut EVA1A-3′UTR in 293 T cells and luciferase assay was conducted. F Hccl-M3 cells were transfected with synthetic mimics of miR-103a-3p (miR-103) or its control miR-NC, inhibitors of miR-103a-3p (anti-miR-103) or its control anti-miR-NC, then the relative expression of miR-103a-3p and EVA1A was determined by RT-qPCR. G, H Hccl-M3 cells were treated as indicated, and the EVA1A protein level was determined by western blot. I Hccl-M3 cells were co-transfected with miR-103a-3p mimics and Myc-EVA1A plasmid, or were co-transfected with miR-103a-3p inhibitors and siRNA of EVA1A (siEVA1A), or transfected with Myc-EVA1A plasmid or siEVA1A. Then the EVA1A protein level was determined by western blot. J Quantitative analysis of the blot bands of EVA1A with ImageJ v1.53 k. Data are shown as means ± SD from three independent experiments. **P < 0.01, ***P < 0.001