Fig. 3
PPARα activation by fenofibrate restores lipid catabolism in Vps15-LKO mice. a Relative transcript levels of metabolic enzymes in ketogenesis, FAO, fatty acid transport and peroxisome biogenesis in livers of fed Vps15f/f and AlbCre+;Vps15f/f mice that were treated for two weeks with fenofibrate. Data are means ± SEM (Vps15f/f (n = 6 and n = 4 for chow and FENO group), AlbCre+;Vps15f/f (n = 5 and n = 4 for chow and FENO group), P < 0.05 *: vs Vps15f/f, #: vs chow, two-tailed, unpaired Student’s t test). b Immunoblot analysis of total protein liver extracts of mice treated as in a using indicated antibodies. Densitometric analyses of protein levels normalised to Pras40 levels presented as folds over Vps15f/f-chow condition. Data are means ± SEM (n = 4 for Vps15f/f chow and FENO group, n = 3 for AlbCre+;Vps15f/f chow and n = 4 for AlbCre+;Vps15f/f FENO group, P < 0.05 *: vs Vps15f/f, #: vs chow, two-tailed, unpaired Student’s t test). c Representative images of immunofluorescent analyses showing nuclear localization of PPARα, NCoR1 and Hdac3 in liver tissue of Vps15f/f and AlbCre+;Vps15f/f mice treated as in a. Secondary anti-mouse or anti-rabbit IgG Alexa Fluor 568 antibody were used for detection. Scale bar: 40 µm. d Relative levels of Acetyl-CoA and Acyl-carnitine metabolites measured by mass spectrometry in liver tissue of mice treated as in a. Data are means ± SEM (Vps15f/f (n = 4 for chow and FENO group), AlbCre+;Vps15f/f (n = 3 and n = 4 for chow and FENO group, P < 0.05 *: vs Vps15f/f, #: vs chow, two-tailed, unpaired Student’s t test). e Immunoblot analyses in total protein liver extracts of 6-week old Vps15f/f and AlbCre+;Vps15f/f mice. Mice were either fed or fasted for 24 h. Four hours prior the sacrifice fasted mice were injected with leupeptin (40 mg/kg) or vehicle. The total protein extracts were immunoblotted with indicated antibodies. f Immunoblot analysis of cytosolic protein fractions of primary hepatocytes that were grown for 72 h in fasting media and treated for 24 h before collection with increasing doses of BafA1. Densitometric analyses of protein levels normalised to Tubulin levels presented as folds over vehicle-treated cells. Data are means ± SEM (n = 4 with 100 nM Bafilomycin A1 treatment, P < 0.05 *: vs vehicle, two-tailed, unpaired Student’s t test). g Relative transcript levels (left panel) of indicated genes in primary hepatocytes incubated in control or fasting media (72 h) treated with or without BafA1 for 24 h before collection. Data are means ± SEM (n = 3, P < 0.05 *: vs control media, #: vs fasting media, two-tailed, unpaired Student’s t test). The immunoblot (right panel) served as control of autophagic activity
