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Cells were subsequently resuspended either in external solution (ES) (containing [mM]: 140 NaCl, 5 KCl, 2 CaCl 2 , 1 MgCl 2 , 10 HEPES, 15 glucose, pH 7.2, 300 mOsm/kg) for transport experiments or in Dulbecco's modified Eagle medium (nutrient mixture F-12 HAM) supplemented with 2 mM glutamine and incubated in a humidified atmosphere with 5% CO 2 and 95% air at 37°C before use in patch-clamp experiments

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Patients are guided through each phase, with close monitoring to adjust interventions as needed