miR-29b/c directly targets DNMT3A.

<p>(A and B) Effects of miR-29b/c on the transcriptional activity of DNMT3A detected by luciferase assay. The 3’UTR of DNMT3A was cloned into the pGL3-basic luciferase reporter vector (DNMT3A 3’UTR-Luc), which was co-transfected with miR-29b/c mimics or negative control mimics (A), or miR-29b/c inhibitors or negative control inhibitors (B) into BGC-823 cells and assayed for luciferase activity. Firefly luciferase values were normalized to Renilla luciferase activity derived from pRL-TK and plotted as relative luciferase activity (*<i>P</i><0.05). The results are expressed as the mean±s.d. of at least three independent experiments. (C and D) qRT-PCR analysis of the DNMT3A relative expression in BGC-823 cells transfected with miR-29b/c mimics (C)/inhibitors (D) or negative control oligonucleotide (*<i>P</i><0.05). (E) Western blot analysis of DNMT3A expression in BGC-823 cells transfected with miR-29b/c mimics (lane 1, 2, 3) /inhibitors (lane 4, 5, 6) or negative control oligonucleotide. <i>β</i>-actin was used as a loading control. The band intensities were quantified and normalized to <i>β</i>-actin intensities with ImageJ software.