Panel A: A bar graph shows the number of cells with informative reads mapped to Tnfrsf9 exons 6/7, 7/8, or 6/8 in different regulatory T-cell subpopulations. The x-axis lists regulatory T-cell subpopulations (Treg-0, Treg-1, Treg-2, Treg-3), and the y-axis shows the Cell number. Different colors and patterns represent different exon splicing events. Panel B: Four Uniform Manifold Approximation and Projection plots display the differentiation pseudotime of Cre-positive Tnfrsf9-positive-positive (wild-type) and Cre-positive Tnfrsf9-flox-flox (knockout) Foxp3-positive regulatory T cells. Each plot is labeled with the lineage (Lineage 1 wild-type, Lineage 2 wild-type, Lineage 1 knockout, Lineage 2 knockout) and shows cells colored by pseudotime. Panel C: Two density plots show the distribution of wild-type or knockout Foxp3-positive regulatory T cells along the pseudotime of Lineage 1 and Lineage 2. The x-axis represents Pseudotime, and the y-axis represents Density. The plots include p-values indicating statistical significance.
Analysis of islet Foxp3 + Treg cluster cells, related to Fig. 5. (A) Analysis of Tnfrsf9 exon 7 (transmembrane domain–encoding exon) alternative splicing in Foxp3+ Tregs of different differentiation states using WT cells shown in Fig. 5 D. The bar plot only contains cells with informative reads mapped to Tnfrsf9 exons 6/7, 7/8, or 6/8. (B) Cre+-Tnfrsf9+/+ (WT) and Cre+-Tnfrsf9fl/fl (KO) Foxp3+ Tregs were combined and analyzed for differentiation pseudotime by Monocle 3. UMAPs of lineages 1 and 2 of Cre+-Tnfrsf9+/+ (WT) and Cre+-Tnfrsf9fl/fl (KO) Foxp3+ Treg clusters are shown. (C) Distribution of WT or KO Foxp3+ Tregs along the pseudotime of lineage 1 or 2. Statistical significance was determined by the Kolmogorov–Smirnov test.