Figure 3

VHL is a direct functional target of miR-566. (A) A 3’ UTR fragment containing the predicted miR-566 targeting sites of VHL were fused downstream of the firefly luciferase gene. (B) Western blot analysis of VHL protein expression in U87 and LN229 cells treated with lenti-NC or lenti-AS-566. (C) U87 and LN229 glioma cells were co-transfected with a human VHL 3’ UTR firefly luciferase reporter plasmid and lenti-NC or lenti-AS-566 as indicated. After 48 h, firefly luciferase activity was measured. (D) Western blot was used to detect the expression of the VHL protein 48 h after transfection of the VHL expression plasmid or control plasmid. (E-H) Overexpression of VHL rescued the effect of miR-566 on glioma cell proliferation (E), cell cycle distribution (F), invasion (G) and apoptosis (H). *P < 0.05.