Analysis of non-ILC immune cell subsets. (A) Sorting strategy for the isolation of Th1, Th2, and Th17 cells. CD4⁺ cells from pooled pLNs and spleen of C57BL/6J mice were magnetically enriched by using anti-CD4 microbeads and the autoMacs Pro separator (Miltenyi Biotec). The enriched CD4⁺ cells were gated on single lymphocytes and sorted in high purity for Th1 (CD3⁺CD4⁺Foxp3⁻CD44^hiCD62L^lowT-bet⁺), Th2 (CD3⁺CD4⁺Foxp3⁻CD44^hiCD62L^lowGata3⁺), and Th17 cells (CD3⁺CD4⁺Foxp3⁻CD44^hiCD62L^lowRORγt⁺). (B) Sorting strategy for the isolation of main immune cell subsets. B cells (CD19⁺), myeloid cells (CD19⁻CD3⁻CD11b⁺), and both CD4⁺ and CD8⁺ T (CD19⁻CD3⁺) cells were sorted from murine lymph nodes. T cells were further distinguished into naive (CD44⁻CD62L⁺) and memory (CD44⁺CD62L⁻) populations. (C) Heatmaps showing the methylation level of ILC1, ILC3, and LTi epigenetic marker regions in Th1, Th2, and Th17 cells. The mean methylation value was calculated from the CpG motifs located within the marker region. The values were translated into a color code ranging from yellow (0% methylation = 0) via white (50% methylation = 0.5) to blue (100% methylation = 1.0).