Adipose tissue SVF isolation and flow cytometry were performed as previously described (46, 47). Briefly, SVF was incubated in Fc Block (rat anti-mouse CD16/32; eBioscience) before staining with anti-CD45 (30-F11), CD3e (145-2C11), CD4 (GK1.5), CD8a (53-6.7), CD11c (N418), and CD64 (X54-5/7.1) (BD Pharmingen). For analysis of Tregs, the samples were fixed after surface staining, permeabilized with a FOXP3 Fix/Perm Buffer Set (421403; BioLegend), and stained with anti-FoxP3 (150D; BioLegend) antibody according to the manufacturer’s protocol. For ILC2 cells, SVF was stained with anti-CD45 (30-F11), B220 (RA3-6B2), CD11b (M1/70), CD11c (N418), Gr-1 (RB6-8C5), CD3e (145-2C11), Thy1.2 (53-2.1) (BioLegend), ST2 (RMST2-2), and Gata3 (TWAJ) (eBioscience), as previously described (11). Samples were analyzed using a BD LSR cell analyzer at the Vision Research Core Facility at the University of Michigan Medical School. Data were analyzed using the FACSDiva v6.2 software (BD Biosciences) and Flowjo (Flowjo.com). Flow cytometry gating strategies for Tregs, B cells, macrophages, neutrophils, CD8+ T cells, and ILC2s are shown in fig. S5.

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