3T3‐L1 adipocyte culture

3T3‐L1 cells were cultured to confluency and treated with preadipocyte expansion media (DMEM + 10% calf serum). After 48 hours, cells were then cultured in differentiation media (DMEM + 10% fetal bovine serum [FBS], 1.0 μM dexamethasone, 0.5 mM 3‐isobutyl‐1‐methylxanthine [IBMX], 1.0‐μg/mL bovine insulin) for 2 days and then cultured in adipocyte maintenance media (DMEM + 10 % FBS, 1.0‐μg/mL bovine insulin) for 10 days. Differentiated 3T3‐L1 adipocyte cultures were then pretreated with DMI (125 or 250 μM) and 4‐OI (250 μM) for 18 hours and then stimulated with lipopolysaccharide (LPS) (100 ng/mL) for 3 hours or palmitic acid (PA, 0.5 mM) or vehicle for 24 hours. The PA:fatty acid free (FFA)‐bovine serum albumin (BSA) conjugate (6:1 molar ratio PA:BSA) was made by dissolving PA in ethanol and then combining with FFA‐BSA dissolved in DMEM. The PA:FFA‐BSA mixture was sonicated to promote solvation of the PA:FFA‐BSA conjugate.