L aspartic acid

BV2 cells were cultured in either DMEM (high glucose; Wako) or DMEM/nutrient mixture F-12 (DMEM/F12; Thermo Fisher Scientific) containing 10% fetal bovine serum (FBS), 100 units penicillin and 100 mg streptomycin (P/S; Thermo Fisher Scientific). For screening, BV2 cells were cultured in DMEM with 10% FBS and P/S in the presence or absence of 150 μM L-alanine (Wako), 50 μM L-glutamic acid (Wako), 50 μM L-aspartic acid (Wako), 50 μM L-asparagine (Wako), 150 μM L-proline (Wako), 0.0052 pM CuSO₄ (Wako), 1.5 pM FeSO₄ (Wako), 1.5 pM ZnSO₄ (Wako), 150 nM linoleic acid (Wako), 500 nM lipoic acid (Sigma), 15 nM biotin (Sigma), 500 nM vitamin B12 (Wako), and 15 μM hypoxanthine (Sigma). Pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid tetrasodium salt (PPADS) was purchased from Alexis Corporation. Dipyridamole (DY) and allopurinol (ALP) were purchased from Tokyo Chemical Industry. Hoechst 33342 was purchased from Thermo Fisher Scientific. DAPI was purchased from Dojindo. FAT3 siRNAs (NM_001080814, siRNA ID; SASI_Mm02_00293873) and MISSION siRNA universal negative control (SIC001) were purchased from Sigma; 50 nM FAT3 siRNAs were transfected using Lipofectamine RNAi Max Reagent (Thermo Fisher Scientific) according to the manufacturer’s instructions.