Figure 4.

miR-15a targets SMAD7. (A) Conserved miR-15a binding site in 3′-UTR of SMAD7. The miR-15a seed match is indicated by vertical lines. (B) Luciferase reporter assays were performed by co-transfection of H9c2 cells with luciferase reporters containing a wild-type or mutant SMAD7 3′UTR sequence together with a control vector or an miR-15a-expression vector. Luciferase activity was normalized to Renilla activity. **P<0.01 vs. control. Representative western blot images and quantification of protein expression levels of (C) SMAD7, (D) cytosolic NF-κB p65 and (E) nuclear NF-κB p65 in H9c2 cells that were non-transfected or were transfected with a NC, miR-15a or anti-miR-15a. Data are presented as the mean ± standard deviation from three independent experiments. *P<0.05 and **P<0.01 vs. H/R. SMAD7, mothers against decapentaplegic homolog 7; miR-15a, microRNA-15a; 3′-UTR, 3′-untranslated region; H/R, hypoxia/reoxygenation; NC, negative control; NF-κB, nuclear factor-κB; Mut, mutant.