Extended Data Fig. 7: scRNA-seq reveals the phenotypic composition of gp33⁻, gp33^int, and gp33^high T cell subsets.

(a) UMAPs of cells profiled by scRNA-seq in the three gp33 T cell fractions colored by the annotated T cell subsets. (b) Venn diagram shows the overlap of all detected TCR clones among the three gp33 T cell fractions. (c) Heat map depicting TCR repertoire overlap (Morisita index) among the different gp33 fractions from the indicated samples. (d) Pie chart representation of the fraction of the detected clone sizes in the three gp33 T cell fractions. (e) Stacked bar plot of the phenotypic distribution of the unique clones from the three gp33 T cell fractions. (f) UMAPs of unique TCR clones determined by scRNA/TCR-seq and colored by the phenotypic distribution of the three gp33 fractions of T cells. (g) Representative flow cytometry plots depict the gating strategy to analyze the fractions of Tex subsets in gp33⁻, gp33^int, and gp33^high CD8⁺ T cells. Bar plots quantify the frequencies of the indicated Tex subsets in each gp33 fraction. Significant changes were determined by two tailed, unpaired t-test at p < 0.05 (n = 3 biologically independent animals). Shown are means with SDs. (h) Representative flow cytometry plots depict the gating strategy to analyze the functionality (IFNG/LAMP1 double positive CD8⁺ T cells) of gp33⁻, gp33^int, and gp33^high CD8⁺ T cells. Boxplot quantifies the double positive fractions of T cells in each gp33 fraction. Significant changes were determined by two tailed, unpaired t-test at p < 0.05 (n = 7 biologically independent animals). Box center line, mean; limits, upper and lower quartiles; whiskers, minimum and maximum values. (i) Stacked bar plots show the phenotypic content of TCR clones that were tested for TCR signaling avidity. CDR3 amino acid sequences are indicated.

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