The expression of effector/exhausted molecules across CD8 ⁺ T cell subsets. (A) Representative plots and cumulative data show the expression of PD-1 and Tox gated on tumor-infiltrating OT-I cells. n = 5 per group. (B) Representative plots and cumulative data show the expression of Tcf1 gated on OT-I cells in TdLNs. n = 4–8 per group. (C) Representative flow cytometry plots and cumulative data show the expression of the indicated molecules gated on tumor-infiltrating CD44⁺ CD8⁺ T cells in the B16F10 tumor model: Mettl8^fl/fl CD4^cre mice and littermate controls were subcutaneously injected with 2 × 10⁵ B16F10 cells. Mice were analyzed at 13 dpi. (D) RNA-seq analysis of Ly108⁺ Tim3⁻ T_PEX, Tim3⁺ CX3CR1⁺ Ly108⁻ Int-T_EX, and Tim3⁺ CX3CR1⁻ Ly108⁻ T_EX cells gated on tumor-infiltrating WT and Mettl8^−/− (KO) OT-I cells. Heatmaps depict stem-like, exhausted and effector-like gene signatures in WT and Mettl8^−/− (KO) OT-I cells. (E) Representative flow cytometry plots and cumulative data show the expression of PD-1 and Tox gated on tumor-infiltrating Tcf1⁺ Tim3⁻ T_PEX, CX3CR1⁺ Tcf1⁻ Int-T_EX, and CX3CR1⁻ Tcf1⁻ T_EX subsets. n = 5 per group. (F) Diamond graphs show chromatin interactions in WT (left) and Tcf1^−/− Lef1^−/− dKO (right) CD8⁺ T cells, with gene structures on the left. (G) Single-cell transcription levels of representative genes illustrated in the UMAP plot. Transcription levels are color coded: gray, not expressed; blue, expressed. (H) Representative flow and cumulative data show the frequencies of CX3CR1, Tim3, and PD-1 in Tcf1⁺ Tox⁻, Tcf1⁻ Tox⁺, and Tcf1⁺ Tox⁺ cells, which are gated from B16F10 tumor-infiltrating CD8⁺ CD44⁺ T cells. n = 9 per group. (I) Schematic diagram of GA treatment to B16F10 cells in vitro (left) and frequency and absolute number of GA-treated cells (right). n = 3 per group. Data are representative of two independent experiments. P value was calculated by two-tailed Student’s t test; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.