Murine Adipocyte Differentiation Assay

Primary MSCs were isolated from the ears of wild-type C57BL/6J, UCP1-KO (Ucp1^tm1Kz/J) (The Jackson Laboratory), and ChREBP KO (kindly gifted by Dr. Lei Yin at the University of Michigan) mice as previously described [39 (link)]. At 2 days post-confluence, adipogenesis was induced with 0.5 mM methylisobutylxanthine, 1 μM dexamethasone, 5 μg/ml insulin, and 5 μM rosiglitazone (Cayman Chemical, Ann Arbor, MI, USA) in DMEM:F12 containing 10% FBS. From day 2 to 4 of differentiation, cells were fed with fresh DMEM:F12 medium containing 10% FBS, 5 μg/ml insulin, and 5 μM rosiglitazone. Thereafter, cells were maintained in DMEM:F12 containing 10% FBS.