Fig. 8: Pharmacological inhibition of ANT heightens naïve T cell responsiveness to activation stimuli.

A Schematic of adoptive transfer: WT mice received daily i.p. injections of CATR or PBS for 14 days. Splenocytes were CellTrace-stained and transferred to WT recipients. Recipients received OVA i.p. one day post-transfer. Three days post-transfer, splenocytes were analyzed for CellTrace intensity and ex vivo restimulation with SIINFEKL peptide. B Representative flow cytometry histograms showing CellTrace intensity gated on donor PBS-treated (gray) or CATR-treated (red) WT OT-I T cells. C Graph depicting proliferation index of donor PBS-treated or CATR-treated WT OT-I T cells (n = 6). D Representative flow cytometry plots showing CD8 vs. IFNγ staining gated on donor PBS-treated (left) or CATR-treated (right) WT OT-I T cells. E Bar graphs summarizing the results shown in D as the frequency of CD8⁺IFNγ⁺ T cells (left), and gMFI of IFNγ gated on CD8⁺IFNγ⁺ T cells (right) (n = 6). F–K Female mice were injected subcutaneously with 1 × 106 B16-OVA tumor cells. Mice were then divided into three groups (n = 5 per group): control (no adoptive transfer), OT-I untreated, and ATR-treated OT-I CD8⁺ T cell adoptive transfer groups. F Intensity of mtDendra2 in naïve OT-I CD8⁺ T cells after spleen dissection. G Representative flow cytometry plots displaying IFNγ versus granzyme B (GzmB) staining in donor untreated and ATR-treated 24-h activated CD8⁺ OT-I T cells. H Bar graphs summarizing the frequency of IFNγ⁺GzmB⁺ (left), and the gMFI of IFNγ⁺ cells (middle), and gMFI of GzmB⁺ cells (right), all gated on donor CD8⁺ OT-I T cells. I Images of tumors from control, OT-I untreated, and ATR-treated OT-I CD8⁺ T cell-injected mice at the end of the experiment (Scale bar: 5 mm). J Tumor volume (mm³) was measured daily day 17 post-tumor implantation. K Tumors were excised, weighed, and presented as tumor weight (mg). Statistical method: two-tailed Mann–Whitney test (C, E, F, H), or two-way ANOVA test (J), or Kruskal–Wallis test (K). Data are represented as mean ± S.D for all expect ±SEM (J). n refers to the number of biologically independent samples. ns = not significant. Source data are provided as a Source Data file.