Figure 8

Generation of 5-hydroxymethylcytosine at specific gene promoters in CD4⁺ T cells. A. - E. The MeDIP approach, using a 5-hydroxymethylcytosine (5hmC) antibody (Active Motif) that specifically recognizes 5-hydroxymethylcytosine, was applied to determine the DNA 5hmC levels at individual CpG dinucleotides located at the promoter regions of inducible genes in CD4⁺ T cells from mice of both age groups during PMA/I stimulation for 0, 0.5, 2 and 4 hours (A. - C.) and T cell differentiation (D. - E.); primers Il2 set B (A) and Il2 set C (B) are located at the Il2 promoter; C . primer Csf2 set A is located at the Csf2 promoter; primer sets A of Ifng and Il4 are located at the Ifng and Il4 promoters, respectively; primers Il17 set A and set B, Foxp3 set A and set B are located at the Il17 and Foxp3 promoters, respectively. The data were normalized to NS, and are shown as the mean and SEM of three independent experiments