Fig 2: ILC2s Are Required for Th2 and Treg Cell Response to IL-33(A–C) WT mice were treated with PBS or IL-33 (i.n., day 0 and 1) and analyzed on day 5 for Foxp3 and GATA3 expression in lung CD4+ T cells (A). Indicated populations (IL-33-treated shown) were subsequently analyzed for expression of Helios and neuropilin-1 (Nrp-1) (B) and CTLA4 (C).(D) RNA-seq was performed on lung Foxp3egfp+GATA3hDC2+ and Foxp3egfp+GATA3hDC2- Treg cells, Foxp3egfp-GATA3hDC2+ Th2 cells, and ILC2s on day 5 after treatment with PBS or IL-33 (i.n., day 0 and 1). Shown is a Venn diagram of transcripts expressed in each cell population (>10 RPKM).(E) Mice were treated with IL-33 and 2W1S-peptide as indicated (i.n., day 0 and 1), followed by quantification of 2W1S-Tetramer+/- Foxp3+GATA3+ Treg cells and Foxp3-GATA3+ Th2 cells in the lung on day 5.(F) Mice were treated with IL-33 (i.n., day 0 and 1) followed by quantification of Foxp3+GATA3+ and - Treg cells and Foxp3-GATA3+ Th2 cells in the lung on day 5.(G) PBS- or diphtheria toxin (DTX)-treated ICOS-T mice were administered with IL-33 and 2W1S-peptide as indicated (i.n., day 0 and 1), followed by quantification of 2W1S-Tetramer+/- Foxp3+GATA3+ Treg and Foxp3-GATA3+ Th2 cells in the mLN on day 5.Bar graphs indicate mean (±SEM). (A)–(C), three repeat experiments, mean percent gated population in (A); (D), single experiment; (E), ANOVA, three repeat experiments; (F), ANOVA, two repeat experiments; (G), ANOVA, three repeat experiments. ns = not significant, *p = 0.05, **p = 0.01, ***p = 0.001, ****p = 0.0001. See also Figures S1 and S2.