For gene expression analysis, animals were euthanized with carbon dioxide from a regulated source. Blood was drawn from the posterior vena cava for clinical chemistry. The liver was weighed and a mid-sagittal section from the left lateral lobe was taken for histology, the remainder of the liver was cubed and frozen in liquid nitrogen. The time from drawing the blood to freezing the liver was less than 90 s. Tissues were stored at -80°C until processing for RNA extraction.

For ATP measurements the animals were anesthetized with pentobarbital (pentobarbital sodium injection, Abbott Laboratories, North Chicago, Il) 50 mg per animal i.p.. In pilot studies we had observed ATP loss in liver samples from animals that were euthanized with carbon dioxide. We used the cold clamp method as described by Easom and Zammit (13).

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