Total cholesterol in undiluted plasma and bile was assessed with Infinity Cholesterol reagent (TR13421, Thermo Scientific, Waltham, MA) and concentrations determined by a standard curve. Liver cholesterol was extracted by homogenizing 50 mg of tissue in a TissueLyser with 1 mL chloroform:isopropanol:IGEPAL CA-630 (7:11:0.1). The organic phase was collected and dried at 50°C. Dried lipids were resuspended in 200μL cholesterol assay buffer (MAK043, Millipore Sigma, St. Louis, MO) and total cholesterol determined following manufacturer’s protocol.
To analyze lipoprotein size distributions, plasma was analyzed using a Superose 6 10-300GL column and size-exclusion fast protein liquid chromatography (FPLC). Fractions were assayed for total cholesterol and triglycerides as previously described[47 (link)].
To analyze lipoprotein size distributions, plasma was analyzed using a Superose 6 10-300GL column and size-exclusion fast protein liquid chromatography (FPLC). Fractions were assayed for total cholesterol and triglycerides as previously described[47 (link)].
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