Extended Data Fig. 3. CD127⁺ ILC1s give rise to CD127⁻ cytotoxic-like ILC1s in vitro and in vivo.

a, b Sorting strategy for CD127⁺ and CD127⁻ liver ILC1 and NK cells from WT mice, gated on live CD45⁺ cells. b Sorting purity. c-f, h, i In vitro cultures of indicated liver ILC1 subsets with IL-2 (c, d, f, h, i) or IL-15 (e) and OP9-DL1 cells (c, e, h, i) or the indicated stromal cell line (d) for 7 days. Representative FACS analysis showing phenotype of NK1.1⁺ cells. Red quadrants on FACS plots highlight phenotypic differences of CD127⁺ ILC1 cultured in the presence of IL-2 versus IL-15. d Frequency of Gzma⁺Gzmb⁺ cells on day 7. e Absolute numbers on d7. Dashed line indicates number of cells on d0 (input=400 cells). f In vitro proliferation of ILC1 subsets assessed by cell tracer violet dilution analyzed on the indicated days. Data are representative of 2 independent experiments with n=4 and 5 (d), n=2 (e), n=4 (f) replicates per experiment. g In vivo co-transfer of congenically marked CD127⁺ and CD127⁻ liver ILC1 into sublethally irradiated Rag2^-/-γc^-/- mice. Representative FACS analysis of liver ILC1 derived from indicated transferred ILC1 subsets on d15. i Isotype control stainings of ILC1 cultured in vitro as in h. Bar graphs indicate replicates (symbols) and mean (bar), error bars display means ± SD.