Zbtb32 augments the short-term immune responses of CD8 ⁺ T cells. (A) Representative FACS plots of TCF1 and Tim-3, CD25, and CD44 in WT and Zbtb32^−/− CD8⁺ cells activated in vitro. (B) Histogram plots (left panel) and MFI (right panel) of IFNγ and GzmB in WT and Zbtb32^−/− CD8⁺ cells activated in vitro. (C) Quantifications of specific molecules in WT and Zbtb32^−/− CD8⁺ cells cultured in vitro (n = 3 in each group). (D) Representative plots of PI and Annexin V in WT and Zbtb32^−/− CD8⁺ cells cultured in vitro.(E) Quantifications of PI and Annexin V in WT and Zbtb32^−/− CD8⁺ cells cultured in vitro (n = 3 in each group). (F) Representative plots and quantifications of CFSE expression level in WT or Zbtb32^−/− CD8⁺ T cells cultured in vitro for 3 days (n = 8 in each group). (G) The quantifications of caspase-3 expression in B16-OVA cells and live cell percentages of activated WT or Zbtb32^−/− CD8⁺ T cells under different E:T ratios in a co-culture killing assay (n = 5 in each group). (H) Expressions of CD25 in activated WT or Zbtb32^−/− CD8⁺ T cells under different E:T ratios in a co-culture killing assay with B16-OVA cells (n = 3 in each group). (I) Schematic representation of the co-transfer of 1.5 × 10⁵ WT and 1.5 × 10⁵Zbtb32^−/− naïve OT-I cells into TCRbd^−/− mice infected with 1 × 10⁵ CFU of LM-OVA (n = 4 in each group). (J) Representative FACS plots of KLRG1 and CD127, IFNγ, and GzmB expressions in WT and Zbtb32^−/− OT-I cells (n = 4 in each group). (K) Quantifications of cell ratio of WT, Zbtb32^−/− OT-I cells, KLRG1 and CD127, and IFNγ⁺GzmB⁺ expressions in WT and Zbtb32^−/− OT-I cells (n = 4 in each group). Data in all graphs are shown as means ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001 by paired or unpaired two-tailed Student’s t test. Data shown are a representative of at least two independent experiments.