Figure 43.

Isolation and analysis of T cells from the murine fat and lung tissue. Gating strategy to identify Treg cells in fat (A) and lung (B) tissue. From all events, a CD4-gate to identify T cells can be drawn (gate G0). Based on G0, lymphocytes can be identified by their FSC/SSC properties (gate G1). Next, doublets are excluded (gate G2) as well as unwanted, dead, or autofluorescent cells (gate G3). From G3, CD4⁺TCRβ⁺ T cells (gate G4) are gated, from which Treg cells (gate G6) and Tconv cells (gate G5) can be identified. Finally, Klrg1⁺ST2⁺ tisTregST2 (gate G7) are gated from Treg cells (gate G6). A staining of Gata-3, where numbers indicate geometric mean fluorescence intensity, exemplifies the expression of this marker in Tconv cells (gate G5, orange, dotted line), Klrg1⁺ST2⁺ tisTregST2 cells (gate G7, red), and Klrg1⁻ST2⁻ Treg cells (gate G8, blue). Numbers indicate frequencies of cells within respective gates. Figures are based on lung and fat digestions from Foxp3^{DTR, GFP} animals.