The concentrations of total cholesterol and triglycerides were determined in whole plasma samples of each mouse with aid of the enzymatic colorimetric assays as described above. The distribution of cholesterol and triglycerides over the different lipoproteins was analysed by fractionation of plasma pools containing equal volumes of all mice in each treatment group using a Superose 6 column (3.2 × 300 mm, Smart-system, Pharmacia). In addition, plasma pools containing samples from 2 mice from per treatment group (total n = 4/5 of pooled samples) were run in an untargeted LC–MS/MS lipidomics analysis as set up by Schoeman et al.33 (link). The metabolite panel consisted of > 100 different species, including essential fatty acids, bile acids, isoprostanes, sphingoid mediators, and lysophosphatidic acids. Integrated peak sizes of each individual metabolite were expressed relative to those of the control group. Plasma alanine aminotransferase (ALT) levels were measured with the ALT activity kit from Sigma according to the protocol of the supplier.
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