Extended Data Fig. 6: ILC1s function independently of DC- and macrophage-expressed IL-15.

a, Schematic describing the IL-15^2A-eGFP reporter mouse strain. b, Types of cancer cells and stromal cells with the potential for IL-15 expression in PyMT tumors. c, Table listing the Cre recombinase lines used to delete IL-15 in the listed target cell populations. d, Schematic of an Il15 floxed allele. e, Gating strategy for determining eGFP expression in the indicated myeloid cell populations isolated from pooled tumors of a 20-week-old IL-15^2A-eGFPPyMT mouse. f, Flow cytometric analysis of eGFP expression in the indicated myeloid cell populations from pooled tumors of a 20-week-old IL-15^2A-eGFPPyMT (colored) or PyMT mouse (gray). g, qPCR analysis of Il15 mRNA expression in sorted DCs from pooled tumors of 20–24-week-old Il15^fl/flPyMT (n = 3) or Itgax-CreIl15^fl/flPyMT (n = 3) mice. h, qPCR analysis of Il15 mRNA expression in sorted TAMs from pooled tumors of 20–24-week-old Il15^fl/flPyMT (n = 3) or Itgax-CreIl15^fl/flPyMT (n = 3) mice. i, Representative plot and quantification of NK1.1⁺ cells out of total CD45⁺CD3⁻ cells isolated from pooled tumors of 20–24-week-old Il15^fl/flPyMT (n = 6) or Itgax-CreIl15^fl/flPyMT mice (n = 6). j, Representative plot and quantification of percentage of CD49a⁺CD103⁺ ILC1s out of total CD45⁺CD3⁻NK1.1⁺ cells isolated from pooled tumors of 20–24-week-old Il15^fl/flPyMT (n = 6) or Itgax-CreIl15^fl/flPyMT (n = 6) mice. k, Representative histogram and quantification of granzyme B (GzmB) expression in CD49a⁺CD103⁺ ILC1s from pooled tumors of 20–24-week-old Il15^fl/flPyMT (n = 5) or Itgax-CreIl15^fl/flPyMT (n = 5) mice. l, (Left) Representative histogram and quantification of granzyme C (GzmC) expression in CD49a⁺CD103⁺ ILC1s from pooled tumors of 20–24-week-old Il15^fl/flPyMT (n = 8) or Itgax-CreIl15^fl/flPyMT (n = 5) mice. m, Total tumor burden of Il15^fl/flPyMT (11 weeks n = 6, 20 weeks n = 11) and Itgax-CreIl15^fl/flPyMT (11 weeks n = 6, 20 weeks n = 11) mice monitored between 11 and 20 weeks of age. n, Representative histograms of CD49b (DX5) expression among total CD3^-NK1.1⁺ cells in spleens of the indicated mouse genotype. (Right) Percentage of DX5⁺ NK cells quantified out of total splenic CD45⁺ immune cells (Il15^fl/flPyMT n = 7, Itgax-CreIl15^fl/flPyMT n = 5). o, (Left) Representative plots of CD27 and CD11b expression among total CD3^-NK1.1⁺DX5⁺ NK cells in spleens of the indicated mouse genotype. (Right) Percentage of DX5⁺CD11b⁺CD27⁻ cells quantified out of total splenic CD45⁺ cells (Il15^fl/flPyMT n = 7, Itgax-CreIl15^fl/flPyMT n = 5). g-o, Each dot represents an individual mouse. Data are pooled from 3 or more independent experiments. All error bars represent the mean ± SEM. Two-tailed unpaired t test was used for statistical analysis, NS = non-significant, *p < 0.05, **p < 0.01, ***p < 0.001.

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