Fig. 4: Fetal lung ILC2s from asthmatic mothers exhibit distinct gene expression compared to the controls.

a Experimental design of snRNA-seq in ILC2s, immune cells, and epithelial cells sorted from fetal lungs of asthmatic or control mothers. b Uniform manifold approximation and projection (UMAP) visualization of snRNA-seq data from all samples. c UMAP of fetal lung ILC2s divided into groups. d Volcano plot showing differences in fetal lung ILC2s between the groups. Differentially expressed genes between OVA and PBS were identified using the FindMarkers function from the Seurat package. The Mann-Whitney U test (two-sided) was used to determine statistical significance, with Benjamini-Hochberg correction applied for multiple comparisons. e Dot plot of representative genes differentially expressed between the groups. Violin plots showing representative cell surface markers and transcription factors differentially expressed between groups (f) and MFI in respective flow cytometry (g). h UMAP depicting subclusters of ILC2s in the fetal lung. i Percentage of ILC2 subclusters compared between groups. Relative expression of Il1rl1, Gata3, and Nr3c1 in ILC2s (j), and a violin plot displaying expression patterns among ILC2 subclusters (k). In (a–k), data were derived from one experiment [(a–f), and (h–k)] with one pregnant dam per group, or two independent experiments (g), each experiment with one pregnant dam per group. Sample sizes were as follows: g PBS: n = 10, OVA: n = 8. In (g), each dot represents an individual mouse. In (g), data are presented as the mean ± SEM. *p < 0.05; **p < 0.01; ****p < 0.0001; ns, not significant [unpaired two-tailed Student’s t-test in (a, h)].