Fig. 1: Ex vivo expanded murine ILC2s exhibit the phenotypic and functional properties of ILC2s.

a Schematic of ex vivo ILC2 expansion. Mice received 4 days of IP IL-25 before the isolation of lineage-negative cells from the peritoneum and mesenteric lymph nodes. Cells were cultured for 6 days in complete media with IL-7 and IL-33. b Scatter plot of H3K4me3 signal from − 300 bp to +500 bp around the TSSs of all transcribed genes and RNA abundance of the corresponding gene (Spearman correlation coefficient, r = 0.81) TPM values represent the average signal of three replicates. H3K4me3 signal represents the average of two replicates. c Distributions of H3K4me3 signal in cells isolated from murine small intestine at TSSs that demonstrated differential H3K4me3 specific to ILC1 or ILC2 cells (n = 2, both conditions, one-sided Mann Whitney U Test, p < = 5.37 × 10⁻⁸). Box plot lines represent min/max values. d Representative tracks of H3K4me3 signal at ILC2 lineage defining genes Gata3, Il13, Il4, Id2, Maf, and Areg. Black bars indicate regions of significant H3K4me3 enrichment (two-sided Wald Test (DESeq2), p adj < 0.05). e, f Following 6 days of expansion, ILC2s were stimulated and cells were characterized by multiparameter flow cytometry. Live, lineage-negative (CD3-, B220-, CD11b-, TER119-, Ly6G-) singlets were assessed for ST2, IFN-y, Tbet, GATA3, and IL-13 expression. e, f Show combined biological replicates where n = minimum of three animals and represent two-three independent experiments. Dotted lines indicate unstimulated controls. Box plots in (c) are centered at the median. The bounds of each box represent the interquartile range IQR, and the whiskers represent the min and max values. Outliers are not shown in boxplots.