Tissues were harvested between noon and 2 PM, 20–24 hours after the last exposure to ethanol or 1 week after the second LPS injection. (DID animals, which were housed on a reverse light cycle, were euthanized during the dark period whereas all other animals were euthanized during the light cycle.) To expedite the procedure and minimize RNA degradation, different individuals performed brain and liver dissections concomitantly. Animals were euthanized by cervical dislocation and brains were removed and placed in a petri dish on ice. After removal of olfactory bulbs, PFC was dissected by cutting the foremost 2 mm of the cortex on each side, at an approximate 50-degree angle from the midline of the brain. Liver samples, approximately 100 mg each, were removed from the lower lobes of the liver. All samples were immediately frozen in liquid nitrogen and stored at −80°C until use.
