FIGURE 4.

Ikaros DNA binding influences DNA methylation at the ifnγ locus. Genomic DNA was purified from wild-type CD4+ naïve (A), wild-type Th2 (B), IkDN/+ Th2 (C), and wild-type Th1 (D) cells and subjected to bisulfite-mediated C > T conversion. Nested primer sets were used to amplify the ifnγ promoter and intron 1 enhancer from converted DNA, and the amplicons were cloned and sequenced. The proportion of the sequenced alleles that were methylated at each CpG site is listed in Table 1 and is depicted in pseudocolor in A–D (blue, 100% methylated; yellow, 0% methylated). The data depicted are derived from 25–27 individual cloned and sequenced alleles. Red boxes highlight regions with differential methylation.