Fig. 6. Effect of HIF-1
Z on HIF-1 activity. Luciferase reporter plasmids containing the Epo enhancer (A) and the VEGF enhancer (B) were transfected into HEK 293 cells transfected with pcDNA and pHIF-1Z. After 16 hours of hypoxia, the luciferase activity of the reporter gene was measured. Each bar, in arbitrary units, represents the mean±s.d. of six experiments. HIF-1 was extracted from the nuclei of the transfected cells subjected to normoxia (N) or 4 hours of hypoxia (H), and analyzed by EMSA (C). For supershift analysis, each 1 µl of rabbit HIF-1 anti-serum, used in Fig. 3B, was added to the completed EMSA reaction mixture. HIF-1 binding (HIF-1), nonspecific binding (NS), and supershifted bands (SS) are indicated. The loading amount of nuclear extract is indicated below the corresponding result. *P<0.05 vs the normoxic control by using unpaired Student’s t-test.
