Figure 4

DNA methylation changes at the promoter of signature genes in T cell differentiation. A. Schematic representation of the promoter regions of Ifng, Il4, Il17 and Foxp3 genes, showing the nucleotide distances upstream from the transcription start sites (dark arrows), the locations of the CpG dinucleotides (vertical arrows) and NF-kB binding elements for reference. PCR primers that covered one or multiple CpG dinucleotides were designed for each inducible gene for the MeDIP assay and are shown as horizontal lines underneath the promoters and denoted by capital letters. B. Using a MeDIP approach, DNA methylation levels at the promoter regions of Ifng and Il4 (B), Il17 and Foxp3 (C) were examined by real-time PCR using the primer sets shown in A. The data were normalized to Gapdh, and were then normalized to the unstimulated samples, and are shown as the mean and SEM of three independent experiments