Fig. 2: Single-cell transcriptome analysis reveals a compositional change of trNK at gd8.5.

a Schematic diagram depicting the procedure of scRNA-Seq library preparation for uterine NK/ILC1. Pooled uteri from five virgin mice or two gd8.5 mice are used. NK/ILC1 cells are sort-purified by a gating strategy of live CD45⁺NK1.1⁺NKp46⁺, followed with labeling by different sample tags and mixed together for subsequent scRNA-Seq library preparation. b UMAP visualization of uterine trNK from virgin and gd8.5 mice. Colors indicate different uterine trNK subgroups. c UMAP of Mki67 expression in uterine trNK (top), and violin plots of Mki67 expression in each uterine trNK subgroup (bottom). d Spearman’s correlation between each uterine trNK subgroup. Whole transcriptomes (left) or transcriptomes with the exclusion of proliferation-related genes (right) are used for the analysis. e Heatmap of the expression of the top 20 specific genes in trNK#1, trNK#2, and trNK#3 subgroups and top 20 proliferation-related genes in the three Mki67⁺ trNK subgroups. f UMAP visualization of uterine trNK in virgin (left) and gd8.5 (right) mice. Colors indicate different uterine trNK subgroups, and ring pie charts show the proportion of each subgroup. g UMAP plot showing expression of Prf1, Gzmb, and Spp1 in uterine trNK from virgin and gd8.5 mice. h Volcano plot showing differentially expressed genes between uterine trNK from virgin and gd8.5 mice. i GO enrichment of the upregulated genes in gd8.5 uterine trNK (h). j Split violin plots showing module score difference between virgin and gd8.5 uterine trNK in each subgroup. The enriched GO terms in (i) was utilized. Data are representative of two independent experiments (b–j).