Female and male ILC2s have cell intrinsic differences.

(A) Amounts of IL-5 and IL-13 produced by purified naïve 8 week old male or female ILC2s stimulated with 5 ng/ml IL-33 and TSLP after 48 h. Red = female, blue = male. Data represented are mean ± SEM, and representative of 2 independent experiments with 4–9 replicates per group. Two-tailed Student’s t-test was used to determine statistical significance, with a P value 5.0 in log2 scale) in at least one sex and were more than 2 fold differentially expressed between male and female. Genes highlighted in red are sex-linked genes. (D) Scatter plot of gene expressions in male and female ILC2s purified from 8-week-old mouse lungs one day after three consecutive injections of IL-33. Gene expressions in log2 scale. Black lines show fold change = 2. Blue = sex-linked genes, red = autosomal genes. Data represented are mean ± SEM, 2 mice per sample, 3 samples per group. (E) Relative expression levels of selected genes in 8-week-old activated male and female mouse lung ILC2s. Genes highlighted in red are sex-linked genes. (F) GSEA of naïve male and female lung ILC2 microarray data in shown in (B). (G) BiNGO analyses comparing naïve female and male mouse lung ILC2 gene expressions. Cytoscape and EnrichmentMap were used for visualizing the functional clusters enriched in each sex and cellular state. Nodes represent enriched GO terms. Node size is proportional to total number of genes in gene set, and node colour to the (1-p value). Edges depict gene set overlap between nodes and thickness represents fraction of shared genes.