Figure S1.
Group 1 ILC–produced TGF-β1 is specifically required for trNK cells in glandular tissues. (A) Expression of Tgfbr2 in group 1 ILCs from indicated tissues in publicly available scRNA-seq datasets (Sparano et al., 2022; Yomogida et al., 2021; Kimmel et al., 2019). (B) Flow cytometric analysis of LAP expression in human peripheral blood mononuclear cells (PBMCs). Tregs were activated with PMA/Ionomycin. FMO, fluorescence minus one. (C) Relative expression of Tgfb1 mRNA in sorted splenic NK cells, T cells and macrophages (Mφ) measured by RT-qPCR. (D) Quantification of key immune cell populations in indicated organs of WT and Ncr1CreTgfb1fl mice. AMs, alveolar macrophages; DCs, dendritic cells. (E and F) Comparison of SG group 1 ILCs (Lin−NK1.1+NKp46+) in WT, Ncr1CreTgfb1fl, and TGF-β receptor 2–deficient (Ncr1CreTgfbr2fl) mice. Representative flow cytometry plots shown in E and quantification in F. (G and H) ILC1 and cNK cell numbers in indicated organs of WT and Ncr1CreTgfb1fl mice. For the liver, helper-like (CD127+IL-18R+) and cytotoxic (IL-18R−GzmB+) ILC1s are shown separately. (I and J) Quantification of BM group 1 ILC numbers (I) and maturation status (J). (B–F and J) Data representative for one of two, or (G–I) pooled from two to three independent experiments with n = 4–8 mice per group. Error bars display means ± SD. Statistical significance was calculated using one-way ANOVA or two-tailed t test; *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. ns, not significant.

Group 1 ILC–produced TGF-β1 is specifically required for trNK cells in glandular tissues. (A) Expression of Tgfbr2 in group 1 ILCs from indicated tissues in publicly available scRNA-seq datasets (Sparano et al., 2022; Yomogida et al., 2021; Kimmel et al., 2019). (B) Flow cytometric analysis of LAP expression in human peripheral blood mononuclear cells (PBMCs). Tregs were activated with PMA/Ionomycin. FMO, fluorescence minus one. (C) Relative expression of Tgfb1 mRNA in sorted splenic NK cells, T cells and macrophages (Mφ) measured by RT-qPCR. (D) Quantification of key immune cell populations in indicated organs of WT and Ncr1CreTgfb1fl mice. AMs, alveolar macrophages; DCs, dendritic cells. (E and F) Comparison of SG group 1 ILCs (Lin−NK1.1+NKp46+) in WT, Ncr1CreTgfb1fl, and TGF-β receptor 2–deficient (Ncr1CreTgfbr2fl) mice. Representative flow cytometry plots shown in E and quantification in F. (G and H) ILC1 and cNK cell numbers in indicated organs of WT and Ncr1CreTgfb1fl mice. For the liver, helper-like (CD127+IL-18R+) and cytotoxic (IL-18R−GzmB+) ILC1s are shown separately. (I and J) Quantification of BM group 1 ILC numbers (I) and maturation status (J). (B–F and J) Data representative for one of two, or (G–I) pooled from two to three independent experiments with n = 4–8 mice per group. Error bars display means ± SD. Statistical significance was calculated using one-way ANOVA or two-tailed t test; *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. ns, not significant.

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