Transferred Tregs exhibit cellular heterogeneity and express a continuum of rTreg- and aTreg-related genes early after transfer but are more “resting-like” by day +20. (A) GSEA enrichment plot of NHP rTreg-specific genes in the ranked list of DE genes between Tregs at day +3 (tSNE cluster C1) compared with transferred Tregs at day +20 (tSNE cluster C2) (top). Dotted circle corresponds to those genes up in rTregs whose expression was higher in day 2 transferred cells compared with day +20. (A) GSEA enrichment plot as in panel A (top), but using NHP aTreg-specific genes (bottom). (B) UMAP dimensionality reduction and clustering of all CFSE⁺ cells and cells from C3 and C4 tSNE clusters that were sorted as endogenous Tregs (left). (B) Cell percentages of sorted cell type (top) and tSNE cluster (bottom) in UMAP clusters from panel A (right). (C) Gene-set scores from panel A plotted in UMAP clusters (left) and UMAP embedding (right). (D) Genes exhibiting high specificity scores for red and cyan clusters.