FIG 7.

EAF2 binding disturbs p300 interaction with HIF-1α. (A) Under normoxic and hypoxic conditions, the enhancement of VEGF promoter activity by cotransfection of HA-p300 was suppressed by overexpression of EAF2 in HEK293T cells (P < 0.0001). (B) Increased levels of EAF2 reduced p300 binding to HIF-1α when the two proteins were overexpressed in HEK293T cells. (C) Overexpression of full-length EAF2 reduced p300 binding to HIF-1α, but overexpression of EAF2 III-VI (which cannot bind to HIF-1α) did not do so. (D) Overexpression of EAF2 in EAF2-null MEFs reduced slc2a expression, but overexpression of EAF2 III-VI did not reduce slc2a expression under normoxia (P = 0.226) and hypoxia (P = 0.0837). (E) Overexpression of EAF2 reduced endogenous p300 binding to endogenous HIF-1α in RCC4 cells. (F) Overexpression of EAF2 reduced HIF-1α-mediated recruitment of p300 to the LDHA promoter, a target of HIF-1α, as revealed by chromatin immunoprecipitation assays (ChIP). Anti-HIF-1α and anti-p300 were used for ChIP assays, and rabbit IgG was used as control. (G) Knockdown of EAF2 enhanced endogenous p300 binding to endogenous HIF-1α in RCC4 cells. (H) Knockdown of EAF2 and FIH-1 enhanced HIF-1α-mediated recruitment of p300 to the PKM2 promoter, a target of HIF-1α, as revealed by chromatin immunoprecipitation assays (ChIP). Anti-HIF-1α and anti-p300 were used for ChIP assays, and rabbit IgG was used as the control.