Fig. 3

Smad4 is a direct miR-34a target. a miRNA target prediction screened one computative miR-34a binding site at Smad4-three prime untranslated region (3′-UTR). b 3′-UTR luciferase reporter assay showed a reduction of relative luciferase activity of wild-type Smad4 3′-UTR by pre-miR-34a in QBC939 and HuCCT1 cells (**P < 0.05). c qRT-PCR analysis of expression of miR-34a treated with miR-34a mimics or miR-34a inhibitor in QBC939 and HuCCT1 cells (***P < 0.01). U6 was used as a loading control. Error bars represent mean ± SD from three independent experiments. d Western blot analysis of Smad4 and Snail expression treated with miR-34a inhibitor or mimic in QBC939 and HuCCT1 cells. β-actin levels were used as internal loading control