Fig. 3: Increased activation potential and effector functions of Ant2^-/- CD8⁺ T cells.

A Representative flow cytometry histograms of CellTrace intensity gated on CD8⁺ T cells derived from WT or Ant2^−/− mice, 72 h post-activation with the indicated concentrations of αCD3 antibody and half the amount of αCD28 antibody. B Bar graph summarizing the results in A as a proliferation index (n = 5). C Representative flow cytometry histograms showing intracellular IFNγ vs. CD25 gated on CD8⁺ T cells derived from WT or Ant2^−/− mice, 24 h post-activation with the indicated concentrations of αCD3 antibody and half amount of αCD28. D–F Bar graphs summarizing the results shown in (C) (WT; n = 6, Ant2^−/−; n = 8), as geometric mean fluorescence intensity (gMFI) of CD25 (D), the frequency of CD8⁺IFNγ⁺ T cells (E), and gMFI of IFNγ gated on CD8⁺IFNγ⁺CD25⁺ (F). G Graph depicts the percentage of killed target B16-OVA melanoma cells after a 6-h incubation at a 1:1 ratio with WT or Ant2^−/− OT-I CD8⁺ T cells activated for 5 days. Target cell viability was assessed using an MTT assay after three washes with PBS (n = 4). H Schematic of the adoptive transfer experiment testing activation of naïve CD8⁺ T cells. CD45.1^+/+ WT recipient mice were intravenously administered CellTrace-labeled CD45.2^+/+ T cells derived from WT or Ant2^−/− OT-1 mice. After one day, recipient mice were intraperitoneally injected with 50 µg of OVA. On day three following cell transfer splenocytes from the mice were examined by flow cytometry for CellTrace intensity. I Representative flow cytometry histograms of CellTrace intensity gated on donor (CD45.2^+/+) CD8⁺ T cells. J Graph depicting the proliferation index of donor WT and Ant2^−/− OT-I T cells (n = 5). Statistical method: Two-tailed Mann–Whitney test. Data are represented as mean ± S.D. n refers to the number of biologically independent samples. ns = not significant. Source data are provided as a Source Data file.