FIGURE 3.

Assessment of hydroxylation reversal on endogenous HIF-1α by immunoblotting with an antibody specific to hydroxy CAD (Asn⁸⁰³). i, representative immunoblot of RCC4 cell lysates probed with antibodies to HIF-1α and hydroxylated HIF-1α (HIF1α N803-OH) following treatment with 1 mm DMOG and cycloheximide. Results are representative of four independent experiments. ii, densitometric analysis of relative band intensities (mean ± S.E., n = 4) of hydroxylated and total HIF-1α protein normalized to the β-actin loading control. Values were plotted with the starting level (t = 0) set to 1.