Fig. 3.

APE1 redox and repair activity are involved in regulating HIF-1 DNA binding to an AP-site containing HRE probe. EMSA of HIF-1 binding with a regular HRE oligo (A) and AP-site-containing HRE oligo (B). Following 2 h of pretreatment with DMSO, myrecitin, E3330 or myrecitin/E3330 at 10 μM for 2 h, HUVECs were cultured in either normal (20%) or hypoxic (1%) conditions for 3 h. The nuclear extracts were prepared for EMSA. HIF-1 antibody was added to the supershift EMSA reaction, whereas ten-fold unlabeled HRE oligos were added to the cold-probe EMSA reaction as an indicator of specificity of the binding between HIF-1 and the HRE oligo. Representative EMSA images are shown, and similar results were obtained in three separate experiments.