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Then samples were centrifuged and washed once with 1 mL of ice-cold 80% methanol.
Cellular uptake was terminated by washing four times with 1 mL of ice-cold PBS.
Then the mixture was washed six times each with 1 ml of ice-cold PBS buffer.
Afterward, 1 mL of ice-cold 1 M sorbitol was added to the cuvette immediately.
The precipitate was twice rinsed with 1 ml of ice-cold acetone at 4°C.
CO was measured in triplets by thermodilution of 10 ml of ice-cold saline.
Radiotracer uptake was stopped by the addition of 1 mL of ice-cold PBS.
Filters were washed three times with 5 mL of ice-cold buffer.
Proteins were precipitated by adding 1 mL of ice-cold acetone and incubating the solution overnight at −20°C.
The resulting partly crystalline residue was precipitated by adding 50 ml of ice-cold water and filtered.
Briefly, dissolved samples were added dropwise to 10 mL of ice-cold methanol (4 °C) with rapid stirring.
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