Extended Data Fig. 5: The locus control region 1 (LCR1), but not LCR2, is specifically required for the development of all tissue resident ILC2s.

a, Representative transcription factor binding motifs enriched in ATAC-seq peaks (peaks from all ILCs pooled) within LCR1 (red section) and LCR2 (blue section). Numbers represent log odds detection scores as computed by HOMER. b, (top) Quantification of absolute numbers of splenic CD4⁺ T cells, CD8⁺ T cells, and NK cells in their different maturation states in LCR1^+/+ and LCR1^−/− mice. (bottom) Quantification of absolute numbers of splenic naive (CD62L⁺, CD44⁻) and effector (CD62L⁻, CD44⁺) CD4⁺ and CD8⁺ T cell populations, as well as CD8⁺ and CD11b⁺ dendritic cells. Splenic T cell numbers are one representative experiment that was repeated six times (n = 8 LCR1^+/+ and 8 LCR1^−/−). Splenic NK cell numbers are a pool of two independent experiments. This experiment was repeated four (n = 9 LCR1^+/+ and 9 LCR1^−/−). Tregs represents a pool of two independent experiments (n = 9 LCR1^+/+ and 9 LCR1^−/−) and was repeated three times. Myeloid cell numbers represent one experiment (n = 3 LCR1^+/+ and 3 LCR1^−/−). Error bars = SEM; and p-values: ns = not significant (Treg panel: Mann-Whitney U two-tailed test; Splenic T cells NK cell and Myeloid cell: Two-way ANOVA with multiple comparison and Bonferroni correction). c, Quantification of absolute numbers of splenic CD4⁺ T cells, CD8⁺ T cells, and NK cells in their different maturation state in LCR2^+/+ and LCR2^−/− mice (n = 4 LCR2^+/+ and 4 LCR2^−/−). Data represents one experiment that was repeated two times. Error bars = SEM; and p-values: ns = not significant, * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.0005 (Two-way ANOVA with multiple comparison and Bonferroni correction). d, Representative flow cytometry gating strategy used for the identification of group 1 (NK cell), group 2 (ILC2s), and group 3 (ILC3s) ILC in small intestine lamina propria (siLP) of LCR1^+/+ and LCR1^−/− animals. e, Quantification of lungs and bone marrow (BM) ILC2s numbers of Arginase1 reporter mouse (Arg1^YFP) and Arg1^YFP; LCR1^−/− mice (Live, CD45⁺, Lineage⁻, CD127⁺, CD90.2⁺, ST2⁺, YFP⁺) at steady state. Data is a pool of two independent experiments and were repeated two times. Each dot represents an individual mouse (n = 7 Arg1^YFP; LCR1^+/+ and 7 Arg1^YFP; LCR1^−/−). Error bars = SEM; and p-values: ns = not significant, * = p ≤ 0.05, ** = p ≤ 0.01, *** = p ≤ 0.0005 (Mann-Whitney U two-tailed test). f, Quantification of IL-5 in the serum of LCR1^+/+ and LCR1^−/− mice at steady state by cytometry bead assay (CBA). Data is representative of two experiments. Each dot represents an individual mouse, (n = 7 LCR1^+/+ and 6 LCR1^−/−). Error bars = SEM; and p-values: ns = not significant, * = p ≤ 0.05 (Mann-Whitney U two-tailed test). g, (top) Representative flow cytometry gating strategy used for the identification eosinophils in the blood, spleen, and lungs of LCR1^+/+ and LCR1^−/− mice at steady state and (bottom) their subsequent quantification. Data represents one experiment. Each dot represents an individual mouse, (n = 4 LCR1^+/+ and 4 LCR1^−/−). Error bars = SEM; and p-values: ns = not significant, * = p ≤ 0.05 (Mann-Whitney U two-tailed test).

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