TGF-βRII deletion results in enhanced transcription of effector-associated genes in more differentiated antigen-specific CD8 T cells. (A) Tim3⁺ CXCR5⁻ differentiated cells were sorted from transferred KO and WT P14 cells in the spleen on D15 and D30 p.i. for RNA-seq. (B) Heatmap showing the relative expression (z-scores derived from expression values) of selected genes in Tim3⁺ KO vs. WT P14 cells on D15 p.i. (C) Volcano plots show log₂ (fold-change) vs. −log₁₀ (adjusted P value) of selected genes upregulated or downregulated in Tim3⁺ KO P14 cells compared with Tim3⁺ WT P14 cells on D30 p.i. The dotted lines indicates log₂ (fold-change) = 0.6 or −0.6, and adjusted P = 0.05. (D) Heatmap showing the relative expression (z-scores derived from expression values) of selected genes in Tim3⁺ KO vs. WT P14 cells on D30 p.i. (E and F) Confirmation of increased expression of IL18Rα (E), and decreased expression of CD200R (F) on Tim3⁺ KO P14 cells by FACS. Paired Student’s t test; *, P < 0.05; **, P < 0.01. (G) GSEA for enrichments of gene signatures of indicated cell types in D15 and D30 Tim3⁺ KO vs. WT P14 cells were visualized by NES and −log₁₀ (FDR q value). RNA-seq datasets were generated from two to three biological replicates for each group at each time point.