Figure 5. HIF-1α prolyl hydroxylation is not required for bcl-2-induced increase of HIF-1α expression and HIF-1 activity in hypoxia.

(A) Western blot analysis of HIF-1α, bcl-2 and PHD2 protein expression and (B) HRE-dependent transcriptional activity in M14 cells stably expressing HA-HIF-1α wild-type (HIF1α wt) or mutated (HIF1α PP/AA), after transiently transfection with control vector (empty), bcl-2 or PHD2 expressing vectors, and then exposure to hypoxia for 24 h. (A) β-actin protein amounts are used to check equal loading and transfer of proteins. Western blot analyses representative of two independent experiments with similar results are shown. (B) Relative luciferase activity of each sample were normalized to the control vector transfected cells. Results represent the mean ± SD of 3 independent experiments performed in triplicate, * p≤0.01.