Adaptable and hardwired features of kidney and SILP ILC2s. (A) Schematic representation of the experimental setup for scRNAseq of ILC2 populations. ILC2s from kidney and SILP were isolated from IL-33–treated BALB/c IL-13^fm mice (i.p. injection on four consecutive days) and sorted for scRNA sequencing analysis (donors). The cells were sorted as CD45⁺Lin⁻CD127⁺IL-13^fm+. Simultaneously, kidney ILC2s from the same isolated cell pool were adoptively transferred i.v. into BALB/c Rag2^−/−Il2rg^−/− mice. After 2 and 8 wk, IL-13^fm+ ILC2s were recovered from SILP and subjected to scRNAseq analysis. (B) Unsupervised UMAP clustering of combined scRNAseq data of kidney and SILP ILC2s from donors and of SILP ILC2s after 2 and 8 wk from recipients. (C) Heat map of differentially expressed ILC2 surface markers in donor and recipient clusters. (D) DE analysis between donor kidney and SILP ILC2s. Genes that are significantly upregulated with ≥twofold change are shown as red dots. (E) Gene set enrichment analysis (GSEA) for HALLMARK pathways of the Mouse Molecular Signatures Database using the genes significantly upregulated in SILP (left, dark blue) and kidney (right, dark green) ILC2s. (F) Violin plots depicting the expression score of SILP program and kidney program, defined as the organ-specific upregulated genes in the DE analyses in D, in the respective conditions. The indicated comparisons represent adaptable SILP features (= comp. 1), non-adaptable SILP features (= comp. 2), and hardwired kidney features (= comp. 3). (G) DE expression analysis of the comparisons indicated in F. Significantly upregulated genes (≥twofold) in the indicated samples are shown as red dots. (H) Venn diagram of SILP-specific and kidney-specific HALLMARK pathways from E and classification of the pathways as indicated. (I) Specific adaptable and non-adaptable HALLMARK pathways in SILP ILC2s identified by filtering as shown in H.