Expression of select transcripts, IR co-expression, and enrichment of key signatures in the scRNA-seq data from CD8 ⁺ TILs from MC38 tumors. (A) Heat map showing Z scores of manually curated genes grouped by biological properties across the eight clusters shown in Fig. 3 B. (B) Co-expression analysis showing distribution of cells expressing 0, 1, 2, 3, 4, or 5 IRs based on transcript levels (transcripts included were Havcr2, Lag3, Tigit, Cd160, and Ctla4.) A cell was determined to be positive for each individual IR if it expressed any count greater than zero. (C) Quantification of IR transcript co-expression shown in B separated by cluster. (D) Enrichment of key gene signatures associated with exhausted CD8⁺ T cell subsets and/or different biological properties (e.g., response to IFN, cell cycle, glycolysis, oxidative phosphorylation). Dot size is proportional to the min-max normalized expression values. Signatures used can be found in Table S2. For the plots in A–D, shown are all cells combined from two independent experiments, representing n = 4 mice per genotype.