Extended Data Fig. 8: DNA methylation regulates the differentiation of ILCPs.

a,b, Representative flow cytometry plots showing the GFP expression in Id2^CreERT2Dnmt1^fl/fl ILCPs (CD127⁺c-Kit⁺α4β7⁺PD-1⁺) transduced with Tbx21-SMR-Snrpn-GFP reporter system in the media without (a) or with (b) 4-OHT. c, d, Bisulfite sequencing analysis showing SMR DNA methylation levels in Id2^CreERT2Dnmt1^fl/fl ILCPs transduced with SMR reporter systems in the media without (c) or with (d) 4-OHT. e, The proportion of the indicated lymphocytes from the thymus, the small intestine (SI), and the lung of Id2^CreERT2 and Id2^CreERT2Dnmt1^fl/fl mice treated as in Fig. 6c. The frequencies of CD45⁺CD4⁻CD8⁻ double negative (DN) T, CD45⁺CD4⁺CD8⁺ double positive (DP) T, CD45⁺CD3⁺CD4⁺ T cells (CD4 T), CD45⁺CD3⁺CD8⁺ T cells (CD8 T), CD45⁺CD3⁺ T cells (T), CD45⁺CD19⁺ B cells (B) were shown as mean ± s.d. (n = 3 or 4 mice per group). f,g, Percentage of cleaved caspase3⁺ ILCPs among total ILCPs (f) and cell numbers of ILCPs (g) from the bone marrow of WT mice, cultured for 2 days with DNMT1 inhibitor (GSK-3484862) or not (vehicle) in vitro. Shown as mean ± s.d. (n = 4 cultures per group). h, i, Percentage of cleaved caspase3⁺ cells among ILC1s, ILC2s or ILC3s (h) and the number of ILC1, ILC2 and ILC3 (i) sorted from the small intestine of WT mice, and cultured for 2 days with DNMT1 inhibitor or not (vehicle) in vitro. Shown as mean ± s.d. (n = 3 cultures per group). Statistical analyses of e-i were performed using two-tailed unpaired Student’s t test. Data are representative of at least three independent experiments.

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