Single-cell RNA-seq reveals Treg subsets and shows that expression profiles of transferred Tregs maintain expression of Treg signatures after adoptive transfer. Visualization of single-cell expression profiles of all sorted cells after tSNE dimensionality reduction colored by unsupervised clustering analysis (A) and sort gate (B, left panel) with yields of sorted cell types in each tSNE cluster (B, right panel). (C) Expression of canonical Treg transcripts and SELL by tSNE cluster. (D) Single-cell scores of a Treg-specific gene-set signature taken from published profiles GSE90600 of human Tregs. (E) GSEA enrichment plot of gene set from panel D among differentially expressed genes between (1) endogenous Tregs and Tconvs (tSNE clusters C5-C10) (green line) and (2) transferred Tregs and Tconv (purple line). (F) C3 and C4 tSNE clusters scored by human aTregs (left) and rTregs (right) gene sets (GSE90600).