SRX10500540 GSM5223077 SRP313121 SRS8624950 GSM5223077 RNA-Seq TRANSCRIPTOMIC cDNA Mice were euthanized and adipose tissue was separated from lymph nodes and surrounding muscle. Isolated tissue was then minced and digested in DMEM (Gibco) containing 2.5% bovine serum albumin, 2 mM L-glutamine, 100 U/mL Penicillin/Streptomycin, 2 mg/mL Collagenase I (Thermo) and 2 mg/mL Collagenase II (Thermo) for 45 min at 37°C with gentle rotation. During the last 15 minutes of incubation 2 mM EDTA was added to the media. Finally, the cell suspension was filtered through a 100 µm strainer. Cells in stromal vascular fraction (SVF) were separated from the adipocyte layer by centrifugation. Single live cells were further purified via fluorescent activated cell sorting, excluding dead cells labelled with LIVE/DEAD Fixable Aqua Dead Cell Stain Kit and doublets based on side and forward light scatter. scRNA-seq of SVF cells was performed using a 10X Genomics Chromium Controller. Single cells were processed with GemCode Single Cell Platform using GemCode Gel Beads, Chip and Library Kits (v2) following the manufacturer's protocol. Libraries were sequenced on HiSeq 3000 (Illumina). Illumina HiSeq 3000 gds 305223077 GSM5223077 GEO Accession GSM5223077 SRX10500541 GSM5223078 SRP313121 SRS8624951 GSM5223078 RNA-Seq TRANSCRIPTOMIC cDNA Mice were euthanized and adipose tissue was separated from lymph nodes and surrounding muscle. Isolated tissue was then minced and digested in DMEM (Gibco) containing 2.5% bovine serum albumin, 2 mM L-glutamine, 100 U/mL Penicillin/Streptomycin, 2 mg/mL Collagenase I (Thermo) and 2 mg/mL Collagenase II (Thermo) for 45 min at 37°C with gentle rotation. During the last 15 minutes of incubation 2 mM EDTA was added to the media. Finally, the cell suspension was filtered through a 100 µm strainer. Cells in stromal vascular fraction (SVF) were separated from the adipocyte layer by centrifugation. Single live cells were further purified via fluorescent activated cell sorting, excluding dead cells labelled with LIVE/DEAD Fixable Aqua Dead Cell Stain Kit and doublets based on side and forward light scatter. scRNA-seq of SVF cells was performed using a 10X Genomics Chromium Controller. Single cells were processed with GemCode Single Cell Platform using GemCode Gel Beads, Chip and Library Kits (v2) following the manufacturer's protocol. Libraries were sequenced on HiSeq 3000 (Illumina). Illumina HiSeq 3000 gds 305223078 GSM5223078 GEO Accession GSM5223078 SRX10500542 GSM5223079 SRP313121 SRS8624952 GSM5223079 RNA-Seq TRANSCRIPTOMIC cDNA Mice were euthanized and adipose tissue was separated from lymph nodes and surrounding muscle. Isolated tissue was then minced and digested in DMEM (Gibco) containing 2.5% bovine serum albumin, 2 mM L-glutamine, 100 U/mL Penicillin/Streptomycin, 2 mg/mL Collagenase I (Thermo) and 2 mg/mL Collagenase II (Thermo) for 45 min at 37°C with gentle rotation. During the last 15 minutes of incubation 2 mM EDTA was added to the media. Finally, the cell suspension was filtered through a 100 µm strainer. Cells in stromal vascular fraction (SVF) were separated from the adipocyte layer by centrifugation. Single live cells were further purified via fluorescent activated cell sorting, excluding dead cells labelled with LIVE/DEAD Fixable Aqua Dead Cell Stain Kit and doublets based on side and forward light scatter. scRNA-seq of SVF cells was performed using a 10X Genomics Chromium Controller. Single cells were processed with GemCode Single Cell Platform using GemCode Gel Beads, Chip and Library Kits (v2) following the manufacturer's protocol. Libraries were sequenced on HiSeq 3000 (Illumina). Illumina HiSeq 3000 gds 305223079 GSM5223079 GEO Accession GSM5223079