Figure 4

MiR-329 suppresses E2F1 expression and Akt pathway. A ) Predicted miR-329 target sequence (blue) in the 3′UTR of E2F1 (E2F1-3′UTR) and positions of the mutated nucleotides (green) in the miR-329 (miR-329 mut). B ) Western blotting analysis of E2F1,pAkt, Akt, p21, cyclinD1 expression in cells transfected with miR-329 or the miR-329 inhibitor, β-actin served as the loading control. C ) Western blotting analysis of GFP expression in the indicated cells, β-actin served as the loading control. D ) Luciferase reporter assay of the indicated cells transfected with the pGL3-E2F1-3′UTR reporter and increasing amounts (10, 50 nM) of miR-329 mimic, miR-329 inhibitor or miR-329 mut oligonucleotides. Bars represent the mean ± SD of three independent experiments. * P <0.05. E ) Luciferase reporter assay of the indicated cells transfected with pGL3-E2F1-3′UTR or pGL3-E2F1-3′UTR-mut reporter with increasing amounts (10, 50 nM) of miR-329 mimic. * P <0.05.