GATA3 mediates the effect of ICAM-1 on ILC2s. (A) Flow cytometric analysis of GATA3 expression in BM iILC2s, pregated on CD45⁺Lin⁻Sca1^highCD127⁺CD25⁺ (n = 10). (B) The mRNA expression of Gata3, Il2ra and Cdkn2b in BM iILC2s (left) or Il5 and Il13 in lung ILC2s (right) from WT and ICAM-1^−/− mice was evaluated by qRT-PCR. (C) BM iILC2s were infected with retrovirus expressing GATA3 or vector control in the presence of 10 ng/ml of IL-2, IL-7, and IL-33 for 3 d. The amount of IL-5 and IL-13 in the culture supernatants was measured by ELISA. (D) Representative flow analysis of the levels of p-Erk1/2 in BM iILC2 from WT (blue) and ICAM-1^−/− (red) after DMSO or PMA treatment for 15 min, and the MFI of p-Erk1/2 were shown. (E) Flow cytometric analysis of GATA3 MFI in BM iILC2s cultured in the presence of IL-2, IL-7, and IL-33 (20 ng/ml) with the indicated treatments for 12 h. (F) The amount of IL-5 and IL-13 in the supernatants of 24 h culture from (E) was measured by ELISA. (G) BM cells from WT and ICAM-1^−/− mice were treated with MG132 (20 µM) and/or U0126 (20 µM) in the presence of 10 ng/ml of IL7 and IL-33 for 6 h. The levels of GATA3 in iILC2 were determined by flow cytometry. (H–J) BM iILC2s from WT and ICAM-1^−/− mice were treated with PBS or LFA-1 protein (0.4 µg/ml) in the presence of IL-2 (10 ng/ml), IL-7 (10 ng/ml), and IL-33 (10 ng/ml). The levels of p-Erk1/2 in ILC2 were measured by flow cytometry after 2 h treatment (H), GATA3 expression in ILC2s was determined by flow cytometry on day 3 (I), and cytokine production by ILC2s was measured by ELISA on day 3 (J). (K) Expression of GATA3 in BM iILC2s after the indicated treatment for 24 h. LFA-1 protein: 0.4 µg/ml; U0126: 20 µM; DMSO: 0.1%. (L) The amount of IL-5 and IL-13 in culture supernatants from K. Data are representative of three independent experiments. Error bars show mean ± SEM; *, P < 0.05; **, P < 0.01; ***, P < 0.001 by unpaired Student's t test (A–D and H–J) or one-way ANOVA with Bonferroni post-test (E–G and K and L). ns, not significant.