ILC2 show partial signature overlaps with Th cells and carry T cell–regulating factor binding sites in overrepresented motifs. (A) B cells (CD19⁺), myeloid cells (CD19⁻CD3⁻CD11b⁺), and naive (CD44⁻CD62L⁺) and memory (CD44⁺CD62L⁻) CD4⁺ and CD8⁺ (CD19⁻CD3⁺) T cells were sorted by flow cytometry from the spleen of WT mice and analyzed for the methylation of CpG motifs within ILC2 marker regions by pyrosequencing. The mean methylation value of all CpG motifs within each region was calculated and transformed into a color-coded box, ranging from blue (100% methylation) and white (50% methylation) to yellow (0% methylation). Each box represents the methylation value of one CpG motif. The white box labeled with nd represents an invalid sequencing signal. The experiment was performed in n = 2 independent sorts. Methylation values shown for the CpG motifs of ILC2 were extracted from the WGBS data of Th1, Th2, and Th17 cells and LN-derived ILC2. B, B cells; M, myeloid cells; Tn, naive T cells; Tmem, memory T cells. (B) Methylation profile of Gata3 gene locus. Smoothed, linear display of CpG motif (bar code) methylation values of the DMR (light gray box) and the surrounding gene body (exons in dark gray boxes, TSS indicated by arrow). Colored lines depict the methylation values ranging from 0 (0% methylation) to 1 (100% methylation) for each Th subset (Th1 = gray, Th2 = green, Th17 = orange). (C) Overrepresented sequences and corresponding E values of identified overrepresented motifs (MEME analysis) among DMRs of ILC2 comparisons are shown as indicated. Motifs containing transcription factors (TFs) not expressed in ILC2 (RNA-seq analysis) or motifs without any transcription factor binding site were excluded. Transcription factors in blue are differentially expressed in ILC2.