Intracellular ATP level was measured using luminescence ATP detection assay (ATPlite, PerkinElmer, USA) according to manufacturer's instructions. Data were reported as arbitrary luminometric units, measured with the microplate reader Wallac Victor multilabel counter and normalized to total protein content.
Evaluation of the glycolytic rate
1.106cells were incubated in 0.5 ml of fresh medium containing 5μCi of [5-3H] glucose (10–20 Ci/mmol; PerkinElmer Life Sciences) for 1 h at 37°C. The reaction was stopped by adding an equal volume of 0.2 N HCl.3H2O was separated from [5-3H] glucose by diffusion in an airtight container for 72 h. Diffused and undiffused tritium was measured using a 1900TR liquid scintillation analyzer (Packard) and compared with controls of [5-3H] glucose only and 3H2O only to determine the rate of glycolysis.
Evaluation of the glycolytic rate
1.106cells were incubated in 0.5 ml of fresh medium containing 5μCi of [5-3H] glucose (10–20 Ci/mmol; PerkinElmer Life Sciences) for 1 h at 37°C. The reaction was stopped by adding an equal volume of 0.2 N HCl.3H2O was separated from [5-3H] glucose by diffusion in an airtight container for 72 h. Diffused and undiffused tritium was measured using a 1900TR liquid scintillation analyzer (Packard) and compared with controls of [5-3H] glucose only and 3H2O only to determine the rate of glycolysis.