Palmitic acid

AML-12 mouse hepatocytes were cultured as instructed. Cells were cultured at 37 °C with 5% CO₂ in a humidified atmosphere. After 24 h serum starvation, cells were exposed to palmitic acid (200 μM, Sigma-Aldrich, USA). For Oil Red O staining, AML-12 cells were cultured in 24-well plates and incubated with palmitic acid (200 μM) for 24 h. AML-12 cells were cultured with PPARα agonist WY14643 (5 μM, MedChemExpress, USA) 30 min before NMDA administration. Then the cells were collected 24 h after the NMDA administration. HepG2 cells were maintained in Dulbecco's Modified Eagle's Medium (HyClone, Thermo Scientific, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Rockville, USA) containing 100 U/mL penicillin and 100 μg/mL streptomycin. HepG2 cells were cultured in 24-well plates and incubated with NMDA (5 mM) for 24 h.

Thermo Acquity UPLC system (Thermo, Waltham, MA, United States) consisted of a photodiode array (PDA)detector, autosampler manager, column compartment, and a binary solvent delivery pump and was connected to Thermo chromeleon software. Inverted photographic microscope was purchased from Tokyo, Japan. Enzyme label detector was purchased from Thermo (United States). palmitic acid was from MedChemExpress (United States). ELISA kits were purchased from BosterBio (United States). Dimethyl sulfoxide (DMSO) was acquired from Invitrogen (United States).