Fig. 3: A_2AR expression is elevated on tumor antigen-specific CD8⁺ T cells within draining lymph nodes but not tumors.

A–G A_2AR eGFP reporter mice were injected subcutaneously with 5 × 10⁵ AT-3 ova tumors. Draining lymph nodes (DLN; A) and tumor infiltrating lymphocytes (B–G) were analyzed by flow cytometry at day 21 post-tumor inoculation. Flow Cytometry plots represent data from concatenated samples in one representative experiment. A Expression of GFP (A_2AR) in CD8⁺ T cells that are CD44⁺Tetramer⁺ or Tetramer^-. Data represent the mean ± SEM of 22 individual mice pooled from 4 experiments. B, C Expression of GFP (A_2AR) on indicated subsets of tumor-infiltrating CD8⁺ T cells. Data represent the mean ± SEM of 17 mice pooled from 3 individual experiments (B) or from 28 mice per group pooled from 5 individual experiments (C). D Mice were treated at days 14, 16, 18, and 20 post-tumor inoculation with 25 µg FTY720. Data represent the mean ± SEM of 5 (Control) or 6 (FTY720) mice per group. E, F Coexpression of A_2AR, PD-1 and CD39 on CD8⁺ T cells (E) and CD4⁺ T cells (F). Data represent the mean ± SEM of 28 (E) or 27 (F) mice per group pooled from 5 independent experiments. G Expression of CD25 on indicated CD4⁺ subsets. Data are concatenated from 6 individual samples. Where normalized MFI is shown, data were normalized relative to the average MFI of total CD8⁺ T cells in each experiment. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Statistics determined by one-way ANOVA (B, C) or unpaired two-sided t test (A, D, E, F). Source data are provided as a Source Data file.