Fig. 5: IL-23 sensing by T_reg cells initiates an eT_reg cell program in the murine TME.

a–h, Foxp3^Cre-YFP/+ (heterozygous) Il23r^fl/fl female mice were inoculated i.d. with B16 tumor cells, and a combined transcriptome (scRNA-seq) and protein expression analysis of sorted CD4⁺ T cells was performed on day 13 after inoculation. Data are shown from one experiment with n = 6. a, UMAP displaying identified tumor-infiltrating T_reg cell clusters (transcriptome; left) and UMAP highlighting Il23-KO and WT T_reg cells (right). b, Heat maps showing adjusted P value (top), average log₂ (fold change) (log₂ (FC); middle) and mean expression (bottom) of subset markers in the identified T_reg cell clusters assessed by Wilcoxon rank-sum test and Benjamini–Hochberg correction. The complete list is available in Supplementary Table 1. c, UMAP displaying identified clusters of integrated tumor-infiltrating and tdLN-derived T_reg cells. d, UMAP of tdLN and tumor T_reg cells with overlayed pseudotime and principal graph lines calculated with Monocle 3. e, Violin plot (left) and UMAP (right) comparing the distribution of WT and Il23r-KO T_reg cells along pseudotime (corresponding to d). f, Neighborhood graph of DA testing results (left). Coloring indicates log (fold change) of differentially abundant neighborhoods between WT and Il23r-KO T_reg cells. White neighborhoods are not differentially abundant (false discovery rate of 10%). Dot size corresponds to the number of cells per neighborhood, and edges indicate the number of overlapping cells between neighborhoods. The index cell position in UMAP space (a) determines ordering of the neighborhood nodes. The Beeswarm plot (right) indicates the distribution of differentially abundant neighborhoods across clustering-based T_reg cell subsets. g, Violin plots depicting the normalized Foxp3 RNA abundance among identified T_reg cell subsets. h, Dot plot displaying selected DEGs between identified T_reg cell subsets encoding transcription factors/signaling proteins (left) and surface or secreted proteins (right). The complete list is available in Supplementary Table 2; cT_reg, central T_reg cells.