FIG. 4.

PHD inhibitor induces CAD activity under normoxic conditions. (A and B) HeLa cells were transfected with GH1α740-826 and pFR-luc reporter (A) or GH1α786-826 and pFR-luc (B). Luciferase assays were performed after the cells were treated with DOG (1 mM), hypoxia (Hpx) (2% O2), and desferrioxamine (Dfx) (130 μM) for 12 h. (C and D) Effects of hydroxylase inhibitor on GH1α740-826 in 786-O#52 (VHL⁺) and 786-O#126 (VHL⁻) cells. (E) Effect of hydroxylase inhibitor on protein stability of GH1α740-826. HeLa cells were transfected with GH1α740-826, pooled, and reseeded in two dishes that were treated with or without DOG. Whole-cell lysates were prepared, and the expression levels of HIF-1α and GH1α740-826 were examined by Western blotting by using anti-HIF-1α and anti-GAL4-DBD monoclonal antibodies, respectively. Cbt, cobalt chloride.