Figure 4. fTreg depletion improves adipose glucose uptake.

(a) Hierarchical clustering of differentially expressed genes between fat Tregs and Tconv and splenic Tregs and Tconv cells from Foxp3-Thy1.1 mice (47 weeks, cells pooled from 3 to 4 mice, same data set used in b,e). (b) FPKM values of selected genes important for Treg identity and canonical suppressive function. (c–d) In vitro suppression assay of fTregs (fTregs pooled from retired breeders, added at 1:1 ratio with splenic conventional T cells, conducted in triplicates). (c) Representative CFSE tracings of conventional T cells with or without fTregs. Gating indicates percentage of dividing cells. (d) Expansion index of Tconv cells. (e) Fold change in expression levels of differentially expressed genes across fat Tregs and Tconv and splenic Tregs and Tconv cells. Fat Treg cluster genes are labeled in red. Position of ST2 is marked. (f) Representative FACS analysis and (g) quantification of ST2 expression in CD4⁺ T cells from aged mice (45 weeks, n=5 mice). (h) Total number of ST2⁺ Tregs and Tconv cells per gram of tissue in VAT and spleen (n=5 mice). (i) FACS histograms and (j) quantification of Tregs (%Foxp3⁺ of CD45⁺ CD4⁺ population) and (k) cells per gram of tissue in VAT and spleen after IL-33 or PBS treatment (16 weeks, n=5 mice per group). (l) Ex vivo glucose uptake in VAT from wild type mice after control or IL-33 treatment (16 week old, n=5 mice per group). (m) Quantification of fTregs and splenic Tregs (%Foxp3⁺ of CD45⁺ population) and (n) ex vivo insulin-stimulated glucose uptake in VAT from wild type mice after anti-ST2 depleting antibody or isotype control treatment (~45 weeks old, n=4 mice per group). (o) Adipo-immune model of metabolic aging. Data represents mean ± s.e.m. *p<0.05, **p<0.01, &p=0.053, %p=0.056.