Gsk2837808a

3T3-L1 preadipocytes were obtained from ATCC (Manassas, VA, USA) and maintained in high glucose-DMEM (HyClone, Logan, UT, USA) supplemented with 10% bovine serum (BS, Gibco 16170, Thermo Fisher Scientific) in a humidified atmosphere of 5% CO₂ at 37 °C. For differentiation, confluent 3T3-L1 preadipocytes were treated with 1 µg/ml insulin (Sigma-Aldrich, St. Louis, MO, USA), 1 µM dexamethasone (Sigma-Aldrich), 0.5 mM IBMX (Sigma-Aldrich), and 10% fetal bovine serum (FBS, Gibco 16000, Thermo Fisher Scientific). After 2 days, the medium was replaced with medium containing 10% FBS and 1 µg/ml insulin and was thereafter replaced every 2 days with medium containing 10% FBS. After 6 days, differentiated 3T3-L1 cells were stained with Oil Red O (Sigma-Aldrich). Oil Red O staining was performed as described previously^{8 (link)}. Sodium dichloroacetate (DCA, 347795), sodium l-lactate (L7022), and chetomin (C9623) were purchased from Sigma-Aldrich. GSK2837808A (# HY-10031) was purchased from MedChemExpress (Monmouth Junction, NJ, USA).

AOAA (C13408), LPS (L2880), Pyruvate (P4562) were purchased from Sigma Aldrich (St Louis, MO, United States). GSK2837808A was purchased from MedChemExpress (New Jersey, United States). Primers were purchased from Sangon Biotech (Shanghai, China).

Western blot assay was performed as previously described [24 (link)]. The PVDF membrane were purchased from Millipore (MA, USA). An informative list of the antibodies was shown in Supplementary Table 3.
MK-2206, 2-DG and GSK2837808A, used in the study, are a chemically synthesized AKT inhibitor, a glucose uptake inhibitor and a lactate dehydrogenase A (LDHA) inhibitor, respectively (all purchased from MedChemExpress, NJ, USA). The working concentration of MK-2206 was 10 μmol/L [25 (link)]. The working concentration of 2-DG was 2 mmol/L [26 ]. The working concentration of GSK2837808A was 10 μmol/L [27 (link)].