Fig. 4.

DFO corrects HIF-1α/p300 binding and augments HIF-1 transactivation and VEGF expression in high glucose culture. (A) HAECs cultured with LG, HG, or HG with DFO (HG/DFO) were exposed to hypoxia and evaluated by mammalian 2-hybrid assay for HIF-1α/p300 binding. DFO treatment augmented HIF-1α/p300 binding in cells exposed to HG. (B) IP/WB of p300 and HIF-1α following treatment described in panel A. β-actin was used as an input control (shown in red typeface). (C) Western blot quantitation of panel B. (D) HIF-1 transactivation analyzed with the 5× hypoxia response element construct described in the Results section. DFO increased HIF-1-mediated luciferase activity in cells exposed to HG in mouse embryonic fibroblasts (MEFs). (E) VEGF ELISA demonstrating DFO's augmentation of VEGF expression by MEFs cultured in HG. *, P < 0.05 vs. HG/DFO group. n = 3.