Fig. 9

In vivo estrogen treatment reduces ILC2 numbers and dampens the IL-33 responses of ILC2. A Flow cytometry gating strategy of ILC2 defined as live, singlet cells lacking lineage markers (LIN: CD11b, CD11c, CD3, CD19, NK1.1, FCeR1, Ter-1) and expressing KLRG-1 and CD127, as well as CD25, ICOS and IL-33R (data not shown for confirmatory markers). B, C Counts of ILC2 in the B BAL and C lungs are shown. D Viability was assessed for ILC2 in bronchoalveolar lavage (BAL) fluid and lungs during flow cytometry analysis. The data are shown as mean ± SEM and are representative of 4 independent experiments. E–H Cytokine expression by lung ILC2 cultured with IL-2, IL-7 and IL-33 for 3 days were determined by ELISA. E IL-5, F IL-13, G CCL3, and H CCL22 are displayed as pg/cell based on ILC2 count determined at the beginning of culture. Results are representative of 3 separate experiments with 4–6 animals included per group. As before, statistical significance was determined using one-way ANOVA with Tukey’s post-test as in previous figures. * Indicates p < 0.05; **p < 0.01, ***p < 0.001, ****p < 0.0001