Fig. 2.

Effect of hypoxia on CpG methylation of early growth response factor-1 (Egr-1) binding site at PKCε promoter and Egr-1 binding in fetal hearts. DNA and nuclear extracts were prepared from 21-day fetal hearts. A: bisulfite-treated DNA was used as a template for real-time methylation-specific PCR. B: competition binding was performed in pooled nuclear extracts with oligonucleotides containing the consensus Egr-1 motifs at −1008 with methylated (M oligo) and unmethylated (UM oligo) CpG. Data are presented as means ± SE; n = 5, analyzed by two-way ANOVA. ^aP < 0.05, hypoxia vs. control; ^bP < 0.05, female vs. male.