Extended Data Fig. 6: CD8⁺ T_EX cells are characterized by high proliferation property and production of effector molecules.

a, A subpopulation of CD8⁺ T_EX shows high expression of MKI67 among 8,530 T cells. b, Gene set enrichment analysis (GSEA) showing the enrichment of proliferation-related pathways in CD8⁺ T_EX cells (n = 3,628; false discovery rate < 0.01; labelled in red). c, Representative example of a CRC tumour stained by multi-coloured IHC showing co-expression of Ki67, CD8, PD-1 and HAVCR2 in CD8⁺ T_EX cells (exemplified by P0413; n = 2 patients). Original magnification, ×20. d, Volcano plot showing the differentially expressed genes between high-proliferative (n = 140) and low-proliferative (n = 720) T_EX cells. Most of the highly expressed genes in high-proliferative T_EX cells are related to cell proliferation. Adjusted P < 0.01; fold change ≥ 2; two-sided unpaired limma-moderated t-test; Benjamini–Hochberg adjusted P value e, Violin plot showing the expression of TBX21, EOMES and PDCD1 in each CD8⁺ T cell (n = 3,628) cluster and the low-proliferative (n = 720) or high-proliferative (n = 140) T_EX cell subsets. f, Most of the clonotypes of high-proliferative T_EX cells were also found in low-proliferative T_EX cells (top). Each row represents an individual clonotype from one patient. Venn diagram showing overlapped clonal clonotypes (≥2 cells) of high- and low-proliferative T_EX cells (bottom). g. Characteristics of CD8⁺ T_EX cells (n = 3,628) as defined by the gene expression of a series of transcription factors, checkpoint receptors, and effector molecules. For violin plots in e and g, colours denote average expression levels; widths denote cell densities.