Lipids were extracted according to the protocol of Bligh and Dyer with modifications (35). Briefly, 0.5 volumes of liver homogenate [1:100 in phosphate-buffered saline (PBS)] were acidified with 0.3 volumes of 0.2 N HCl. Subsequently, 3 volumes of MeOH/CHCl3 (2:1, v/v), 1 volume of CHCl3, and 1 volume of H2O were added and mixed vigorously stepwise. The phases were separated by centrifugation, and the lower phase was transferred to a new tube and evaporated under a stream of nitrogen. The lipids were either resuspended in DMSO/H2O for enzymatic quantification of TG and cholesterol (as described before) or in MeOH/CHCl3 (1:1, v/v) for separation by TLC on a silica gel 60 plate (Merck) in the solvent mixtures of CHCl3/MeOH/20% acetic acid (65:25:5, v/v/v), hexane/ethyl acetate/acetic acid (59:10:1, v/v/v), and pure hexane. Cholesteryl palmitate (for CE), triolein (for TG), oleic acid (for FFA), cholesterol (for FC), phosphatidylethanolamine (for PE), and phosphatidylcholine (for PL) were used as standards for the TLC. Lipid spots were visualized by dipping the plate in a hydrous solution of 2.5% 12-molybdophosphoric acid (Alfa Aesar), cerium (IV) sulfate (Sigma), and 6% H2SO4 and heating at 200°C until the bands appear. Densitometric analysis was performed using ImageJ and known concentrations of the standards.

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