Figure 2. TCR specificity was a critical driver of VAT Treg accumulation.

(A, B) Proliferation and decay rate of clonotype⁻ (non-Vα2⁺Vβ4⁺) and clonotype⁺ (Vα2⁺Vβ4⁺) VAT Treg cells. A) Frequencies of Ki67⁺ VAT Treg cells from 10wk-old Tg⁺ male mice (n=6). Data for Vα2⁺Vβ4⁺ VAT Tregs also appear in Fig. 3E. B) 10wk-old Tg⁺ male mice were given BrdU in the drinking water for 4 weeks, and BrdU⁺ VAT Treg cells were detected at various time points after drug removal (n≥3).

(C–E) CD4⁺ T cells from pooled spleen and LNs of Tg⁻ or Tg⁺ CD45.2⁺ male mice were iv-transferred into B6.CD45.1⁺ male mice. C) Frequencies and numbers of CD45.2⁺ Treg cells at 12wks after transfer (n=5). D) Frequencies of Vα2⁺Vβ4⁺ cells in CD45.2⁺ TCR-tg Treg cells at 12wks after transfer (n=5). E) Time course analysis following transfer of TCR-tg CD4⁺ T cells (n=5).

(F) Treg cells sorted from Tg⁻ and Tg⁺ CD45.2⁺ Foxp3-GFP^KI/y male mice were transferred into B6.CD45.1⁺ male mice. Frequencies and numbers of CD45.2⁺ Treg cells at 12wks after transfer (n≥7).

(G) PPARγ (above) and ST2 (below) expression in Tconv cells from 20wk-old Tg⁻ and Tg⁺ Pparg-Tdt^KI/+Foxp3-GFP^KI/y male mice (n≥6).

(H, I) Experimental set-up as per panels C–E. H) Frequencies of CD45.2⁺ cells in Tconv cells at 12wks after transfer (n=5). I) Time course analysis following transfer of TCR-tg CD4⁺ T cells (n=5).

(J, K) Tconv cells sorted from Tg⁻ and Tg⁺ CD45.2⁺ Foxp3-GFP^KI/y male mice were transferred into B6.CD45.1⁺ male mice. J) Frequencies of CD45.2⁺ cells in CD4⁺ T cells at 12wks after transfer (n=5). K) Frequencies of Treg cells converted from CD45.2⁺ Tconv cells at 12wks after transfer (n=5).

Summary plots show data pooled from two to four independent experiments. Mean ± SD.