Fig. 6.

Functional characterization of intratumoral CD8⁺ T cells. (A) GSEA of RNA-seq data for cell cycling-related gene transcripts in cells sorted from orthotopic MMTV-PyMT tumors at day 5 posttreatment (4 mice per cell type per treatment). Positive and negative scores indicate increased and decreased cell cycling activity, respectively. (B) Flow cytometry analysis of Ki67 of adoptively transferred CD45.2⁺ OVA-CD8⁺ T cells in MC38-OVA tumors (Left) and TDLN (Right). Each data point represents one mouse. (C and D) Quantification of perforin⁺ cells in MC38 tumors. (C) Data indicate the number of perforin⁺ cells per unit of area. Each data point represents one tumor, of which at least 6 images were analyzed. (D) Representative images show antiperforin (brown) immunostaining and hematoxylin/eosin staining (light blue) of tumor sections. (Scale bar, 50 μm.) (E) RNA sequencing of Prf1 gene expression in CD8⁺ T cells sorted from orthotopic MMTV-PyMT tumors at day 5 posttreatment (4 mice per treatment). Data are shown as log2-transformed RPKM (reads per kilobase per million mapped reads). Each data point represents one mouse. Data indicate mean values ± SEM. Statistical analyses by 1-way ANOVA with Tukey’s correction for multiple comparisons (black) or pairwise Student’s t test (red) unless otherwise indicated in Dataset S2. The number of mice employed in each experiment is reported in Dataset S2.