scRNA-seq analysis of T cell subsets and their heterogeneity in tumors of different treatment groups. Mice were challenged with B16-F10 cells and were given indicated treatments; tumors were isolated and digested. TILs from tumors were isolated and stained with anti-CD45.2 antibody for sorting by FACS, and a 10X library was prepared and analyzed as described in the Materials and methods. The CD3⁺ cells were reclustered, and T cell subpopulations were characterized. (A) UMAP graph showing the clusters and annotation. (B) UMAP graphs showing the expression of selected markers. (C) UMAP graph showing the clusters in each treatment group. (D) Bar plot of the frequency of each T cell cluster. Cluster names are indicated on the y axis and frequencies on the x axis. Data represent two independent experiments (n = 5 mice per group). Mice within each group were pooled for analysis. Irradiated parental B16 tumor cells (Vac) were used as a control for the IVAX group.