Fig. 6

Smad4 is a direct downstream target of miR-34a (n = 3). a The target sites of miR-34a in the 3′-UTR of Smad4 were shown as a schematic representation predicted by TargetScan. The normal seed sequence and mutant were highlighted and underlined. b Luciferase activity with wild-type or mutant Smad4 3′-UTR was performed after co-transfection with the miR-34a mimic or inhibitor or respective controls into HEK293T cells. c and d The levels of Smad4 mRNA and protein expression in SMMC-7721 cells with knocked-down miR-34a expression or its negative control, and in HCC-LM3 cells overexpressing miR-34a or its negative control using western blot and qRT-PCR. NC, negative control. WT, wild-type. *P < 0.05; **P < 0.01