Subcutaneous adipose tissue and VAT samples were either frozen at −80°C if unfractionated, or immediately processed if they were to be separated into adipocyte and SVF. Unfractionated fat (~200 mg) or muscle (100 mg) were mixed with zirconium oxide beads and 1.0 mL of TRIzol (Thermo-Fisher) in a 1.6 ml microcentrifuge tube (Thermo-Fisher) and treated with a tissue disruptor (Bullet Blender Storm 5, Next Advance) at full speed for 2 min. RNA was purified from the homogenized samples using the RNAeasy Lipid Tissue Mini kit (Qiagen). Nucleic acid (~1.0 μg) was next treated with DNase 1 (Promega). After DNase inactivation, RNA was reverse transcribed using the RNA to cDNA high capacity kit (Thermo-Fisher), following manufacturer protocol.