No mice died in any group after 3 weeks of feeding with the control or ethanol-containing diet. Before being sacrificed, the mice were fasted overnight and body weight was measured. Blood was collected and serum was separated. Whole liver was removed and liver weight measured; the liver was rapidly excised into fragments and washed with cold saline, and one aliquot of tissue was placed in 10% formalin solution for paraffin blocking. Another piece was placed in Tissue-Tek O.C.T Compound, and then was frozen and cut into 10-micron frozen sections for oil red O staining. and for DHE fluorescence assays. The remaining aliquots were stored at −80°C for further assays. Liver homogenates were prepared in ice-cold 0.15 M KCl. All samples were stored at −80°C in aliquots.
