Figure 2.

Analysis of tumor-infiltrating immune cells in CT26.WT tumors. CT26.WT cells were inoculated (1 × 10⁶ (a–c) or 6 × 10⁵ (d–f) cells per mouse) into the right flank of BALB/c mice. After tumors were established, mice were treated with ASP7657 twice per day. (a–c) Modification of the myeloid cell population. Tumors were excised from mice treated for 10 days, enzymatically dissociated, stained, and analyzed using flow cytometry (n = 6 per group). Frequencies of DCs (a, CD45⁺ CD11b⁺ CD11c⁺ MHC-II^high 7-AAD⁻ cells in CD45⁺ 7-AAD⁻ cells), M-MDSCs (b, CD45⁺ CD11b⁺ Ly6G^low Ly6C^high 7-AAD⁻ cells in CD45⁺ CD11b⁺ 7-AAD⁻ cells), and M1-TAMs (c, CD45⁺ CD11b⁺ F4/80⁺ MHC-II^high 7-AAD⁻ cells in CD45⁺ CD11b⁺ 7-AAD⁻ cells) are plotted. Modification of the T cell population. Tumors were excised from mice treated for 14 days, enzymatically dissociated, stained, and analyzed using flow cytometry (n = 9 − 10 per group). Frequencies of CD8⁺ T cells (d, CD45⁺ CD3⁺ CD4⁻ CD8⁺ 7-AAD⁻ cells), CD4⁺ T cells (e, CD45⁺ CD3⁺ CD4⁺ CD8⁻ 7-AAD⁻ cells), and conventional T cells (f, CD45⁺ CD3⁺ 7-AAD⁻ cells) in 7-AAD⁻ cells are plotted. Veh: vehicle; ASP: ASP7657 3 mg/kg/day. Data show mean ± SEM. ^∗p < 0.05 compared to the vehicle-treated group (Student's t-test).