FIGURE 4.
Degradation of HIF-1α(P402A/P563A) is not achieved by overexpression of PHDs 1-3. A-C, PHD1, PHD2, and PHD3 fail to induce degradation of HIF-1α(P402A/P563A). HEK 293 cells were transfected with pFLAG-mHIF-1α, or pFLAG-mHIF-1α(P402A/P563A) (PP-A) and 150 ng (+), or 300 ng (++) of plasmids encoding FLAG-tagged PHD1 (A), FLAG-tagged-PHD2 (B), or FLAG-tagged-PHD3 (C). Whole cell extracts were separated by SDS-PAGE and analyzed by immunoblotting using anti-FLAG (α-FLAG), and anti-actin (α-Actin) antibodies. Cells were treated with CoCl2 as indicated (+).