Nutrient depletion and rescue of GOT1-null 143B cells with different metabolites

To deplete nutrients in the medium, both wild type and GOT1-null 143B cells were seeded in 96-well plates in triplicate at a density of 5 × 10³. The cells were continuously grown for 4 days without changing medium. Then the medium was replaced with glucose-free and 1 g/L glucose DMEM supplemented with 10 mM aspartate, 5 mM OAA, 2.5 mM PEP respectively. For glucose-free DMEM, malate (10 mM), succinate (10 mM) and NAD⁺ (100 μM) were also tested. The cell morphological changes were recorded. The cell viability treated with malate, succinate and NAD⁺ for 24 h were determined with XTT assy.