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Fig. 3 | Genome Medicine

Fig. 3

From: Single-cell transcriptomics reveal that PD-1 mediates immune tolerance by regulating proliferation of regulatory T cells

Fig. 3

Coreceptor and costimulation blockade-induced transplant tolerance is predominantly mediated by intragraft proliferation of CD4+ Treg. a Biaxial scatter plots by t-SNE analysis showing single-cell transcriptomic clustering of ~ 1000 CD4+hCD2 (TH) or ~ 1000 CD4+hCD2+ (TR) cells purified from rejecting and tolerated grafts, respectively. Cells were colored individually according to their initial expression of CD4 and hCD2 during FACS sorting. b t-SNE analysis showing single-cell transcriptomic clustering of ~ 1000 CD4+hCD2+ Treg cells during transplant rejection and tolerance, respectively. Cells were further subgrouped into specific clusters (C1-4) and colored individually according to expression patterns of specific marker genes and spatial proximity in the biaxial plot. c From selected pathways determined by GO functional annotations in terms of biological processes of each cluster (Additional file 1: Tables S4–S6), upregulated genes with an average expression level > 0.05 were displayed by the heatmap. d Scran analysis showing cell cycle phase classifications. e Immunostaining and f quantifications of Ki67+FOXP3+ cells among total FOXP3+ cells in rejecting and tolerated grafts. Inserts denote magnified Ki67+FOXP3+ cells. Scale bars in e: 50 μm and inserts: 10 μm. n = 6 per group, **P < 0.01. Abbreviations: R-TH, CD4+hCD2 Th of rejecting grafts; R-TR, CD4+hCD2+ Treg of rejecting grafts; T-TH, CD4+hCD2 Th of tolerated grafts; and T-TR, CD4+hCD2+ Treg of tolerated grafts

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