Characterization of TCR diversity and clonotype expansion by scVDJ-seq. (A) Clonal expansion mapped onto the UMAP from Figure 1B. (B) Shannon diversity index estimated from TCR sequencing results for 45 random cells per cluster. Values are shown for each cluster identified in Figure 1B. (C) Histograms showing the percentages of Treg subsets with expanded clonotypes within the entire Treg compartment and across samples of tonsils (n = 3), FL (n = 3), and DLBCL (n = 3). Expanded clonotypes were categorized as small expansion (1 < X ≤ 3) and medium expansion (3 < X ≤ 6). (D) Distribution of CDR3 length for each Treg cluster as per the tissue origin. The mean CDR3 lengths for activated Tregs (actTregs), resting Tregs (restTregs), and LAG3⁺ Tregs were 28.1 amino acids (aas), 28.3 aas, and 27.9 aas, respectively in FL; 28.0 aas, 27.7 aas, and 28.3 aas in DLBCL; and 28.3 aas, 28.0 aas, and 28.1 aas in tonsils. (E) Venn diagrams showing the number of clonotypes within and shared between Treg subsets. For scaling purposes, numbers are shown for the Tregs found among 1000 random CD4⁺ T cells with gene expression and single TCRα/TCRβ gene.