Mouse 3t3 l1 cell lines

Mouse 3T3-L1 cell lines were obtained from the American Type Culture Collection. The cell lines were cultured in Dulbecco modified Eagle medium (DMEM, Gibco™, Thermo Fisher Scientific, Waltham, MA, USA) with 10% fetal bovine serum (FBS, Gibco™, Thermo Fisher Scientific) at 37 °C in a 5% CO₂ incubator. Adipogenic differentiation was carried out according to the classical MDI (1-methyl-3-isobutylxanthine, dexamethasone and insulin) cocktail. 3T3-L1 preadipocytes were grown to confluence. Two days later (designated as D0), 3T3-L1 preadipocytes were stimulated for two days in differentiation medium: DMEM containing 10% FBS and MDI (0.5 mm 3-isobutyl-1-methylxanthine, 1 μm dexamethasone, and 5 μg/mL insulin). Two days later, the medium was changed to DMEM containing 10% FBS and 5 μg/mL insulin, and the medium was refreshed every two to three days until harvest.

Mouse 3T3-L1 cell lines were purchased from American Type Culture Collection (ATCC), (Manassas, VA, USA). The cell culture reagents including Advanced Dulbecco’s modified Eagle’s medium (DMEM), GlutaMAX, Penicillin-Streptomycin (10,000 U/mL), and fetal bovine serum (FBS) were ordered from Gibco Life Technologies (Thermo Fisher Scientific, Waltham, MA, USA). The Oil Red O solution, 3-isobutyl-1-methylxanthine (IBMX), dexamethasone, Rosiglitazone, and insulin were obtained from Sigma-Aldrich (St. Louis, MO, USA). PureLink RNA purification kit by Ambion, cDNA synthesis kit, TaqMan Array Mouse GPCR Panel, advanced TaqMan PCR Master Mix, and SYBR Green PCR Master Mix were purchased from Applied Biosystem (Thermo Fisher Scientific, Waltham, MA, USA). DNA oligonucleotides were obtained from Macrogen. Antibodies were obtained from Thermo Fisher (Thermo Fisher Scientific, Waltham, MA, USA) and Cell Signaling Technology (Danvers, MA, USA).