Extended Data Fig. 6: Effector differentiation of ILC1s is regulated by Hobit across tissues.

a Gating strategy identifying ILC1s among live CD45⁺ cells in the small intestine lamina propria for data depicted in Fig. 7a. To preserve the Hobit^Tom signal, cells were gated without transcription factor staining. ILC1 (1) were enriched as Lin⁻ NKp46⁺ NK1.1^hiCD90^int CXCR6⁺ CD127⁺ cells, NK cells (2) were gated as Lin⁻ NKp46⁺ NK1.1^hi CD90^int CXCR6⁻ CD127⁻ cells, NKp46⁺ ILC3 were gated as Lin⁻ NKp46⁺ CD90^hi cells. Purity of gating strategy is displayed by analysis of RORγt and Eomes (right). b-q Analysis of ILC1 in salivary glands (b-e), kidneys (f-i), mesenteric lymph nodes (j-m) and small intestine lamina propria (n-q). b, f, j, n Representative FACS analysis of Lin⁻NK1.1⁺ cells (left row of FACS plots) and Eomes⁻CD49a⁺ ILC1s (middle and right row) of WT and Hobit^KO mice. b Representative FACS plot identifying ILC1 (Eomes⁻CD49a⁺), Eomes⁺CD49a⁺NK1.1⁺ cells (Eomes⁺CD49a⁺) and cNK cells (Eomes⁺CD49a⁻) within Lin⁻NK1.1⁺ cells (left panel). c, g, k, o Bar graphs indicate absolute numbers of ILC1. d, h, l, p Frequency of ILC1s expressing indicated marker proteins. e, i, m, q gMFI of CD127, TCF-1 and IL-18R1 expression within ILC1 that are gated as positive for the respective marker. Data are representative of 3 independent experiments with n=3 (c, e), n=4 (j, l, m, p, q) and n=5 (b, d, f-i, k, o) mice per group. Bar graphs indicate individual mice (symbols) and mean (bar), error bars display means ± s.d. Statistical significance was calculated by unpaired two-tailed t-test; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

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