B cells and NK cells are dispensable for the terminal differentiation of Sirpα⁺CD4⁺Esam⁺ cDC2s. (A) Representative flow cytometry plots and quantification of cDCs in the spleens of WT and uMT^−/− mice. n = 7 WT mice, and 7 uMT^−/− mice; two independent experiments; two-way ANOVA with Sidak’s multiple comparisons post-test. (B) Gating strategy for the identification and quantification of NK cells and ILCs. NK cells are defined as Lineage(CD3e/CD19/Ter119)⁻NK1.1^hiNKp46^hi cells. ILCs are identified as Lineage(CD3e/CD19/Ter119)⁻NK1.1^−/lowNKp46^−/lowCD90⁺ cells. The different ILC subsets are further defined based on the expression of IL7Ra, Rorgt, NKp46, Tbet, Gata3, and ST2. (C) Quantification of innate lymphocytes in the spleen of mice treated with depleting NK1.1 or CD90 antibodies. n = 12 Rag2^−/− + isotype antibody control mice, 11 Rag2^−/− + aNK1.1 mice, and 8 Rag2^−/− + aCD90 mice; at least two independent experiments per treatment regimen; Kruskal–Wallis test with Dunn’s multiple comparisons post-test. (D) Representative flow cytometry plots of selected innate lymphocyte populations and quantification of innate lymphocytes and cDC in the spleen of WT and NKp46-DTA mice. n = 4 WT mice, and 4 NKp46-DTA mice for ILC quantification; one experiment; Mann–Whitney U test. n = 8 WT mice, 7 littermate WT control mice, and 8 NKp46-DTA mice for cDC quantification; two independent experiments; two-way ANOVA with Sidak’s multiple comparisons post-test. (E) Quantification of splenic Sirpα⁺CD4⁺Esam⁺ cDC2s in the spleens of Rag2^−/−γc^−/− mice following adoptive transfer of ILCs. n = 11 Rag2^−/− x γc ^−/− PBS control mice, 13 Rag2^−/− x γc^−/− + ILCs; three independent experiments. Results are discretized by the recovery rates of Tbet⁺ ILC1s, Gata3⁺ ILC2s, and Rorgt⁺ ILC3s, with the number of samples in each group varying according to the number of ILCs of each type recovered. Kruskal–Wallis test with Dunn’s multiple comparisons post-test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.