Fig. 4

miR-663a directly targeted TGF-β1. a The overlap of TGF-β1 by bioinformatics prediction between miRWalk, TargetScan, PITA, DIANAmT and miRanda programs. b Diagram of the putative binding site of miR-663a on the 3′UTR of TGF-β1 predicted by Targetsan. The mutant sequences of 3′-UTR of TGF-β1 used in luciferase reporter is shown in transverse line. c Relative expression of luciferase reporters with wild type TGF-β1 3′-UTR or mutant TGF-β1 3′-UTR after co-transfection with miR-663a agomir, antagomir or respective control in 293 T. d qRT-PCR analysis of TGF-β1 mRNA expression in the HCC cells transfected by either miR-663a agomir, antagomir or respective control. The expression of TGF-β1 was normalized against GAPDH. e Western blot analysis of TGF-β1 protein expression in the HCC cells transfected by either miR-663a agomir, antagomir or respective control. The expression of TGF-β1 was normalized against Tubulin. Data are presented as mean ± SD (n = 3). *p < 0.05, **p < 0.01 and ***p < 0.001