RA signaling drives functional adaptation of ILC2s to the intestinal phenotype. (A) Dot plot of RA receptor expression in the scRNAseq data set of kidney and SILP ILC2s from donors and of SILP ILC2s after 2 and 8 wk from recipients of kidney ILC2s (see Fig. 4 A for experimental setup). (B) Heat map of RA receptor expression in bulk RNAseq data set of ILC2s isolated from the kidneys, lungs, and SILP of naïve C57BL/6 mice (see Fig. 1). (C) Dot plot of RA signaling genes selected from the DE analyses (see Fig. 4 D) and top 30 cluster-defining genes (see Fig. S4 A). (D) Heat map of RA signaling genes in the bulk RNAseq data set of ILC2s isolated from naïve C57BL/6 mice (see Fig. 1). (E) Violin plots depicting the expression score of RA signaling associated genes in the respective conditions. (F) Color mapping of RA expression score on the UMAP subclustering of adapting ex-kidney ILC2s at 2 wk after transfer (see Fig. 5). Lowest expression is indicated by blue and highest expression by yellow. (G) RNA velocity-based pseudotime analyses of selected RNA signaling genes in adapting ex-kidney ILC2s at 2 wk after transfer. (H and I) Representative histogram overlays (H) and MFI (I) for surface marker expression of sorted kidney ILC2s after culture with IL-2 (n = 4 for each condition) in the presence or absence of RA (1 µM). (J) Cytokine quantification in the supernatant of the cultured kidney ILC2s stimulated with subthreshold IL-25 or IL-33 (1 ng/ml each; n = 4 for each condition). Data in H–J are representative of three individual experiments with similar results. Symbols represent individual data points and bars indicate mean ± SEM. Statistical analysis was performed using unpaired two-tailed Student’s t test (*P < 0.05, **P < 0.01, ***P < 0.001). (K) Hypothetical model of RA signaling network in SILP ILC2s based on genes upregulated in the RNAseq data sets and a literature review of described downstream pathways. CREB, cAMP response element-binding protein; ISG, interferon-stimulated gene.