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Cells were washed three times with Krebs buffer (20 mM HEPES, 5 mM KH 2 PO 4 , 1 mM MgSO 4 , 1 mM CaCl 2 , 136 mM NaCl, 4.7 mM KCl, pH 7.4), then incubated with 500 g/ml 2-NBDG, a fluorescence-labeled deoxyglucose analog, for 20 min at room temperature