Figure 2

Transfection of synthetic miR-200 and luciferase reporter gene assay. (A) qPCR and western blot of VASH2 in HepG2 cells transfected with miR200a/b/c mimics and inhibitors (*represents p < 0.05 compared with the control group). (B) Bioinformatics analysis predicted the binding sites of miR200a/b/c in the VASH2 3’UTR (R1-R5 represent 5 separate binding sites). (C) VASH2 3’UTR luciferase reporter assay assessing the post-transcriptional regulation of VASH2 by miR200a/b/c (*represents p < 0.05 compared with the mutant group; R1-R5 represents wild-type binding sites; R1M-R5M represents mutant binding sites). miR200a/b/c significantly inhibited luciferase activity in constructs containing the wild-type R1, R2 and R3 binding sites compared with the mutant binding sites, but no effect was observed for constructs containing R4 and R5.