Figure 2. Dynamics of tumor-infiltrating CD4⁺ T cell subsets in mouse breast cancer models.

A. and C. Proportions of CD4⁺ T cell subsets in TILs were analyzed at the indicated time points using flow cytometry analyses by gating CD4⁺ population. The treatment procedure was identical as that described in Figure 1. TILs were isolated and intracellular staining performed after stimulation with PMA and ionomycin for 5 hours. B. and D. Relative cell numbers of CD4⁺ T cell subsets with the tumor progression were calculated based on their absolute cell numbers per tumor volume in each tumor tissue. E. Dynamic changes of Foxp3⁺CD4⁺ to CD8⁺ T cell ratios were analyzed based on the proportions of CD4⁺Foxp3⁺ subset and CD8⁺ TILs. Result of each dot shown in A. to E. is derived from an individual mouse. Data shown are mean ± SE from four mice in each time point. *p < 0.05 and **p < 0.01 between the indicated two groups determined by paired student's t test. Data shown in A. to E. are representative from three independent experiments with similar results.