2.16. Glycerol assay for lipolysis

Mouse serum was used to measure glycerol concentration as per manufacturer's instructions. For cell culture, 3T3-L1 preadipocytes were differentiated for 7 days in a 12-well plate following a standard protocol [32]. Lipolysis was measured following a standard procedure [29]. Briefly, adipocytes were incubated with DMSO or TNP (1 μM) for 4 h in DMEM containing 0.5% BSA (fatty acid free). Afterwards, media was changed, and fresh media was added with or without isoproterenol (IsoP; 10 μM). TNP treatment was continued throughout the assay. Cell culture media was used for glycerol assay using the same kit described above.