Figure 3.

64B prevents the binding of p300 and CBP to HIF-1α. A, Western blot of indicated protein expression after dose-response of 64B in Mel290-V6R cells after 24 h of hypoxia. B, Western blot of indicated protein expression after time-course of hypoxia +/− 64B (2 μmol/L) treatment. C, D, E, Co-immunoprecipitation (co-IP) of cell extracts from Mel290-V6R and 92.1 UM cells grown for 24 h. under normoxia or hypoxia +/− 64B (2 μmol/L) treatment. Note reduction in binding between HIF-1α and p300/CBP cofactors, but not HIF-1α. Western blot analysis of an aliquot (4%) of the cell extracts was used as a control for input protein (input). F, Western blot analysis of indicated protein expression in 92.1 cells under normoxia and hypoxia +/− FIH sliencing. G, Effect of siRNA-mediated silencing of FIH, on 64B’s ability to inhibit p300 - HIF-1α interaction in hypoxic 92.1 cells as measured by co-IP.