Figure 3.

Luciferase reporter assays with reporter constructs containing (A) HIF-1α (B) OSGIN2 and (C) VEGF 3′UTRs. Relative luminescence coefficients were standardized on a scrambled mimic transfected control. Co-transfection with 20 nM miR-199a-5p yielded a significant downregulation of the luciferase-coupled HIF-1α-3′UTR construct (35.4%, P=0.041, Student's t-test) and the OSGIN2-3′UTR construct (26.2%; P=0.029, Student's t-test, error bars defining standard derivation) compared with controls (n=3; *P<0.05). No such downregulation was observed in VEGF 3′UTRs. (D) Western blot analyses presented a downregulation of HIF-1α and OSGIN2 expression following miR-199a-5p (20 nM) overexpression. HIF-1α, hypoxia-inducible factor 1α OSGIN2, oxidative stress induced growth inhibitor 2; VEGF, vascular endothelial growth factor; UTR, untranslated region; scRNA, scrambled RNA; RD, Rhabdomyosarcoma cells.