Notch1 and Jagged1 are directly regulated by miR-34a.

<p>(A) Putative binding sites of miR-34a in Notch1, Notch2 and Jagge1 3′UTR are shown with color letters. (B) Jagged1, Notch1, Notch2, Snail protein level and mRNA level were respectively detected by Western blotting and qRT-PCR 48 h after transfection with miR-34a precursor or precursor control in HK2 cells under low oxygen. A representative blot from three independent experiments is shown (left). The histogram shows the average volume density normalized to the β-actin (right). (C) Dual luciferase assay was performed in HK2 cells under hypoxia co-transfected with luciferase construct (pMIR-Norch1-3′UTR, pMIR-Norch2-3′UTR, pMIR-Jagged1-3′UTR or pMIR-report control) and miR-34a precursors or precursor control. Firefly luciferase activity was normalized to Renilla luciferase activity for each sample. The results shown represent the mean ± SD from 3 independent experiments. <i>^#p<0.05 and *p<0.01</i> compared with empty vector or miRNA precursor control.