Fig. 3: Liver CD8⁺ Trm cells from DKO mice exhibit enhanced activation and increased cytotoxicity.

a Heatmap of mRNA expression level of TCR signaling, cytokine production and cytotoxicity-associated genes of CD8⁺ Tcm, Tem and Trm cells from liver of DKO mice. b Representative FACS histogram of Granzyme A (GzmA), Granzyme B (GzmB), Perforin, FasL and CD107a expression on CD8⁺ Tcm, Tem and Trm cell subsets. The ranged gate and the percentages of the positive population are shown in the figure. c Heatmap of mRNA expression level of co-inhibitory moleculars associated genes of CD8⁺ Tcm, Tem and Trm cells from liver of DKO mice. d Representative FACS histogram of selected immune-checkpoint molecule expression on CD8⁺ Tcm, Tem and Trm cell subsets. The ranged gate and the percentages of the positive population are shown in the figure. e UMAP plots of DKO liver CD8⁺ Trm cells, colored by cell clusters. f Violin plots showing mRNA level of Cd244a and Cd200r1 of different Trm subset. g Differentiation trajectory of Trm subset by monocle analysis. Blue arrow indicating the probable direction of differentiation. h Representative flow plots of CD200R and 2B4 expression on CD8⁺ Trm cells. i Heatmap of mRNA expression level of exhaustion, stemness, cytotoxicity and costimulatory molecule associated genes of CD8⁺ Trm subsets. j Representative FACS plot of Trm clusters and representative FACS histogram graphs of TCF-1, CD127, PD-1, Annexin V, IFN-γ and Perforin expression on Trm subsets. The ranged gate, the percentages of the positive population and gMFI of Perforin are shown in the figure. k Representative flow plots of Annexin V and DAPI in PIBECs co-cultured with corresponding CD8⁺ T cell subsets. l Percentage of hepatic DAPI⁺ or Annexin V⁺ cells in PIBECs cocultured with corresponding CD8⁺ T cell subset from DKO mice liver. n = 3 cells examined over three independent experiments. Data in (b, d, h, j) are representative results of at least three independent experiments. The p values were determined by a one-way ANOVA with Tukey’s multiple comparisons test (l). Data are shown as Means ± SEM.