Fig. 3

Increased miR29b levels reduce PI3K/AKT pathway components and DNMT3B expression. U266 and RPMI 8226 cells were transfected with 10 µM miRNA mimic negative control (miR-NC) or miR29b mimic (miR29b) for 72 h. A Levels of miR29b in transfected cells normalized to RNU44 and expressed as fold change relative to the average value for untransfected cells (real-time PCR and 2^−ΔΔCt method). B Representative western blots of PI3K/AKT pathway components. C Densitometric analysis of the experiment shown in (B). Band intensities were normalized to β-actin and then to values for untransfected cells (dotted line). D Levels of DNMT3B mRNA in transfected cells, normalized to GAPDH and expressed as fold change relative to the average value for untransfected cells (real-time PCR and 2^−ΔΔCt method). E Representative western blot and densitometric analysis of DNMT3B in transfected cells. Values were normalized first to β-actin and then to untransfected cells. Values are mean and SD for 7 independent experiments. *P < .05. **P ≤ .01 Wilcoxon signed-rank test