Characterization of SG group 1 ILCs in adult mice. (A) Immunofluorescence staining showing EpCAM (dark blue), CD31 (magenta), and NKp46-TdTomato (yellow) in indicated organs of Ncr1^CreR26R^Ai14 (WT) and Ncr1^CreR26R^Ai14Tgfb1^fl mice. Scale bars, 20 μm. (B and C) Fluorescently labeled anti-CD45 antibody was injected intravenously (i.v.) to differentiate intravascular and extravascular leukocytes. (B) Flow cytometric quantification of indicated cell populations in adult mice showing percentage of intravascular (CD45i.v.⁺) cells. (C) Representative flow cytometry plots of SG group 1 ILCs at indicated age. (D) Eomes and CD62L-based flow cytometric gating strategy for SG cNK cells and corresponding expression of CD49a in WT and Ncr1^CreTgfb1^fl cNK cells. (E–G) Publicly available scRNA-seq data of adult SG group 1 ILCs (Yomogida et al., 2021) were reanalyzed and compared to flow cytometric data in Fig. 3. UMAP with clusters (E) based on subset-defining marker expression shown as heatmap (F). (G) Color-coded expression of indicated genes. (H and I) Flow cytometry gating strategy to differentiate SG ILC1s from trNK cells (H) and representative plot showing expression of GzmB and GzmC (I). (J) Histograms showing expression of indicated markers in SG group 1 ILC subsets. (A) Data representative for n = 10 mice, or (B–D and H–J) representative for one of two to three independent experiments with n = 3–6 mice per timepoint or group. Error bars display means ± SD. Statistical significance was calculated using one-way ANOVA or two-tailed t test; ****P < 0.0001. ns, not significant.