Supplementary Figure 6 : BATF and IRF4 are required for VAT-Treg cell development.

From: The transcriptional regulators IRF4, BATF and IL-33 orchestrate development and maintenance of adipose tissue–resident regulatory T cells

Supplementary Figure 6

(a-b) Proportion of Treg cells in the spleen and VAT of wild-type (WT) mice compared to Batf–/– (b) and Irf4–/– (b) mice. Values are means ± S.D. from 5-7 mice per group. (c-d) VAT mass (c) and body weight (d) of WT, Irf4–/– and Batf–/– mice. Values are the means from each 6-8 mice per group (one way ANOVA). (e) Bar graph showing proportions of WT and knock-out Foxp3+ cells as indicated from the spleens and VAT of Ly5.1 (WT) / Batf–/– (left) and Ly5.1 (WT) / Irf4–/– peripheral chimeric mice. (f) Flow cytometric analysis of ST2 expression on Treg cells from the VAT of mice of the indicated genotype. (g-h) MACS enriched CD4+CD25+ cells from WT (Ly5.1), Batf–/– (Ly5.2) and Irf4–/– (Ly5.2) mice as indicated were mixed as indicated, CTV labeled and cultured in conditions that induce ST2. Flow cytometric analysis of total Foxp3+ cells. Numbers indicate percentages of cells. Bar graphs show the proportion of Foxp3+ cells of the indicated genotype that express ST2. Histograms (gated on Foxp3+ cells) show CTV dilution profiles. Values are mean ± S.D. from 5 male 30-week-old mice per group. (i) RNAseq track showing expression of GzmB by cTreg cells and eTreg cells. (j) Weight of VAT from Irf4fl/flGzmBCre+ and control mice. Values are mean from 4 mice per group. *P<0.01; **P<0.002; ***P=0.0001; ****P<0.0001 (unpaired, two tailed t-test)..