Fig. 5

RT plus YM101 promoted CD8⁺ T cell activation in the 4T1 murine model. (a-h) Flow cytometry analysis of tumors from BALB/c mice subcutaneously inoculated with 4T1 cells and sacrificed two days after the completion of treatment. Quantification of total T cells (a), CD8⁺ T cells (b), activated CD8⁺ T cells (CD69⁺) (c), Ki67⁺ CD8⁺ T cells (d), TNF-α⁺ CD8⁺ T cells (e), IFN-γ⁺ CD8⁺ T cells (f), Perforin⁺ CD8⁺ T (g) and Granzyme-B⁺ CD8⁺ T cells (h). Each point represents an individual mouse (n = 5 per group). (i) Representative images from CD8 (red), CD4 (green) and FOXP3 (yellow) in CT26 tumors stained by IF. Scale bars represent 50 μm. (j) Quantification of CD8⁺ T cell density (cells/mm²) in multiplexed IF images of tumor area. (k) CD8⁺ T cell/Treg ratio in tumor area based on multiplexed IF analysis. (l-n) Effect of CD8⁺ T cell depletion on combined therapy in the 4T1 tumor model. (l) Tumor volume expressed as the mean ± SEM. Data are from two independent experiments with 6 or 9 mice per group in each experiment. (m) Tumor weights on day 12 (n = 6 per group). (n) Kaplan-Meier plot of survival by treatment group and median survival are presented. The P values were determined using a one-way ANOVA followed by Dunnett’s multiple comparisons test. *P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001