Figure S1.
Characterization of ILC subsets and ILC2s in different organs in BALB/c and C57BL/6 mice. (A) Flow cytometric characterization and gating strategy of ILC subsets isolated from kidney, lung, and SILP of naïve BALB/c mice (blue = ILC2s, red = ILC3s, and yellow = ILC1s). Numbers indicate the percentage of events in the respective gates. (B) Frequencies and absolute numbers of total ILCs in kidney, lung, and SILP. (C) Frequencies of ILC subset distribution in the respective organs. Symbols represent individual data points, and bars indicate mean ± SEM. (D) Flow cytometric analysis of Lin−GATA-3+ ILC2s from PBS- or IL-33–treated (i.p. injection on four consecutive days) wild type BALB/c mice at day 12–14 after the first injection. Heat maps show MFI of various surface markers of ILC2s in the indicated organs normalized to surface marker expression of kidney ILC2s. (E) Unbiased UMAP clustering of the flow cytometry data from D. Plots show combined and single-organ contribution of kidney (orange), lung (blue), and SILP (green) ILC2s to the UMAP clustering. Data are representative of at least two independent experiments with similar results with n = 3–5 mice per group. (F) Representative histogram overlays showing surface marker expression on Lin−GATA-3+ ILC2s isolated from kidney (orange), lung (blue), and SILP (green) of PBS- or IL-33–treated (i.p. injection on four consecutive days) wild type C57BL/6 mice. (G) Concatenated flow cytometry data from A shown in an unbiased UMAP clustering along with multigraph color mapping of cell surface markers on UMAP projection. Lowest expression is indicated by blue and highest expression by red. (H) Absolute numbers of different cytokine-producing ILC2s (see Fig. 1, L and M) isolated from the kidney, lung, and SILP of PBS- (n = 6) or IL-33–treated (n = 5) wild type C57BL/6 mice. Data are pooled from two individually performed experiments.

Characterization of ILC subsets and ILC2s in different organs in BALB/c and C57BL/6 mice. (A) Flow cytometric characterization and gating strategy of ILC subsets isolated from kidney, lung, and SILP of naïve BALB/c mice (blue = ILC2s, red = ILC3s, and yellow = ILC1s). Numbers indicate the percentage of events in the respective gates. (B) Frequencies and absolute numbers of total ILCs in kidney, lung, and SILP. (C) Frequencies of ILC subset distribution in the respective organs. Symbols represent individual data points, and bars indicate mean ± SEM. (D) Flow cytometric analysis of Lin−GATA-3+ ILC2s from PBS- or IL-33–treated (i.p. injection on four consecutive days) wild type BALB/c mice at day 12–14 after the first injection. Heat maps show MFI of various surface markers of ILC2s in the indicated organs normalized to surface marker expression of kidney ILC2s. (E) Unbiased UMAP clustering of the flow cytometry data from D. Plots show combined and single-organ contribution of kidney (orange), lung (blue), and SILP (green) ILC2s to the UMAP clustering. Data are representative of at least two independent experiments with similar results with n = 3–5 mice per group. (F) Representative histogram overlays showing surface marker expression on Lin−GATA-3+ ILC2s isolated from kidney (orange), lung (blue), and SILP (green) of PBS- or IL-33–treated (i.p. injection on four consecutive days) wild type C57BL/6 mice. (G) Concatenated flow cytometry data from A shown in an unbiased UMAP clustering along with multigraph color mapping of cell surface markers on UMAP projection. Lowest expression is indicated by blue and highest expression by red. (H) Absolute numbers of different cytokine-producing ILC2s (see Fig. 1, L and M) isolated from the kidney, lung, and SILP of PBS- (n = 6) or IL-33–treated (n = 5) wild type C57BL/6 mice. Data are pooled from two individually performed experiments.

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