Figure 4. Suppression of ILC2 function by Ahr is cell-intrinsic.

(A to E) Sorted small intestinal (SI) ILC2s (Lin⁻KLRG1⁺CD90⁺) from WT (Ahr^+/+) (CD45.1/CD45.1) or KO (Ahr^−/−) (CD45.2/CD45.2) age and sex-matched mice were mixed equally (20,000 cells in total) and transferred into Rag2^−/−Il2rg^−/− littermate recipient mice. Experimental design (A), and FACS analyses of CD45.1 and GATA3 expression (upper), ST2 and GATA3 expression (middle), and IL-5 and IL-13 expression (bottom) after gating on indicated populations (B). Data are representative of two independent experiments. Percentages of ST2⁺ (C), IL-5⁺ (D), and IL-13⁺ (E) cells in recovered WT or KO ILC2s (Lin⁻GATA3⁺). Data are shown as mean ± SEM (n=3 per group). (F to K) RFP⁺ ILC2s (Lin⁻ KLRG1⁺RFP⁺) were sorted from Ahr^f/f Il5^RFP-Cre or littermate Ahr^+/+ Il5^RFP-Cre mice. FACS analyses of Ahr expression (F), ST2 and GATA3 expression (G), and IL-5 and IL-13 expression (I) after gating on sorted RFP⁺ ILC2. Data are representative of two independent experiments. Percentages of ST2⁺ (H), IL-5⁺ (J), and IL-13⁺ (K) cells in sorted RFP⁺ ILC2s (Lin⁻GATA3⁺). Data are shown as mean ± SEM (n=3 per group). (L and M) Deletion of Ahr in vitro by retroviral expression of Cre-recombinase in ILC2s. Experimental design (L), and FACS analyses of Thy1.1 and SSC (left) after gating on ILC2s (Lin⁻GATA3⁺), and ST2, IL-5, IL-13, and Ahr expression after gating on Thy1.1⁺ (Cre⁺) or Thy1.1⁻ (Cre⁻) ILC2s (M). Data are representative of two independent experiments. See also Figures S6 and S7.