Figure 2. IL18rl⁺ST2^- ILCs in neonatal and adult lung share similarities with IL18rl⁺ BM ILCPs.

(A-C), t-SNE maps of neonatal lung ILC single-cell transcriptomes highlighting RaceID3 clusters (A) or sorted ST2^- and ST2⁺ ILCs (B), and log2 normalized expression of representative genes marking putative ILC progenitors (C). (D) Expression of candidate genes within neonatal lung clusters with at least 10 cells. Color represents z-score mean expression across clusters and dot size represents fraction of cells in the cluster expressing the gene. (E) FACS analysis of neonatal (P4) and adult (6-8w) Il18r1⁺ST2^- lung ILCs, represented as t-SNE maps, indicating expression levels for Gata3, RORγt, PLZF, Tcf7, C-kit, CD103 and Ki67. (F) Histograms comparing ILC subsets (PLZF⁺Gata3⁺, RORγt⁺CD103^-, RORγt⁺CD103⁺, Il18r1^-ST2^- ILC2s and RORγt⁺ ILC3) in adult lung for Gata3, Icos, a4p7, PD1, CD25 and Il17rb expression levels. Data are representative of 2-3 independent experiments (n=6-12 mice). (G) t-SNE map of Il18r1⁺Icos⁺ BM ILCs highlighting RaceID3 clusters. (H) Expression of candidate genes for Il18r1⁺Icos⁺ BM clusters with at least 10 cells. Color represents z-score mean expression across clusters and dot size represents fraction of cells in the cluster expressing the gene. (I-J) FACS analysis of human lung and PB ILCs pre-gated on CD45⁺Lin^- CD127⁺EOMES^-T-bet^-CRTH2^-NKp44^-c-KIT^+/- (I) or c-KIT ILCs (J). Figures depict representative staining (I) and t-SNE maps (J) indicating the expression levels of selected markers. Data are representative of 3 independent experiments (n=2-4 samples each).