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ATP extraction was performed as previously described [ 70], with minor modifications.
ATP standard curve construction and ATP extraction from oocysts and sporozoites was performed using a freeze/thaw lysis procedure previously described [39], [40].
ATP concentration was normalized on mass of liver slices used for one ATP extraction.
ATP was extracted from cells by the addition of 50 μl of ATP extraction reagent (DCS) to each well of the 96-well plate.
The method for cAMP extraction using TCA was modified from a protocol for ATP extraction (Gustafsson, 1979).
Reproducible results were obtained during ATP extraction with 8 10 mg of S. cerevisiae cells using 1 ml of boiling ethanol.
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The drug concentrations used are given in table 3. The plates were incubated for six days at 37°C with 5% CO2, and the remaining ATP extracted with tumor cell extraction reagent (TCER; DCS).
The plates were incubated for 6 days at 37°C with 5% CO2, following which ATP was extracted with tumour cell extraction reagent (DCS).
The aldose reductase pathway can therefore preserve endoneurial ATP when oxygen extraction is limited by capillary dysfunction, in that it reduces the ATP expenditure needed to maintain energy-efficient oxidative phosphorylation.
Extraction of ATP, ADP and AMP were conducted with the method of Ozogul et al. [ 60] with a minor modification.
To facilitate the extraction of ATP, 0.5 mL Extractant B/S (BioThema, Sweden) was added to each tube and vortexed for 5 seconds.
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