Hepatic Lipid Profiling and Plasma Analysis

Approximately 100 mg liver was homogenized in methanol and lipids were extracted in chloroform/methanol (2:1 v/v) as described ^{43 (link)}. Hepatic triglyceride and cholesterol levels were then quantified using Infinity reagents from Thermo Scientific (Waltham, MA). Plasma lipid and glucose levels were also determined using Infinity reagents. Plasma lipoprotein profile was analyzed by FPLC as described ^{19 (link)}. Briefly, after 100 μl plasma was injected, lipoproteins were run at 0.5 ml/min in a buffer containing 0.15 M NaCl, 0.01 M Na₂HPO4, 0.1 mM EDTA, pH 7.5, and separated on a Superose 6 10/300 GL column (GE Healthcare) by using BioLogic DuoFlow QuadTec 10 System (Bio-Rad, CA). 500 μl of sample per fraction was collected.