Fig. 6
Fenofibrate treatment rescues mitochondrial function in Vps15-LKO mice. a Representative liver transmission electron micrographs of control or fenofibrate treated Vps15f/f and AlbCre+;Vps15f/f mice. The fold change of relative mitochondria area and mitochondria number are presented as graph. Data are means ± SEM of calculations of total ~1500 mitochondria normalized to total area of the fields analysed (n = 15–20 randomly taken fields, n = 2–4 mice, P < 0.05 *: vs Vps15f/f, #: vs chow, two-tailed, unpaired Student’s t test). Scale bar: 1 µm. Protein (b) and relative transcript levels (c) of mitochondria fusion factor, mitochondrial DNA transcription factors, mitochondrial DNA polymerase and respiratory chain subunits in liver of Vps15f/f and AlbCre+;Vps15f/f mice treated with fenofibrate. Densitometric analyses of protein levels normalised to eIF2a levels presented as folds over Vps15f/f-chow condition. Data are means ± SEM (n = 4–6 for Vps15f/f chow and FENO group, n = 4–5 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). d Primary hepatocytes were transduced with adenoviral vectors expressing PGC1α, PGC1α + Vps15 or GFP protein as a control and 24 h post-transduction were transfected with reporter construct. The relative luminescence presented as fold difference over empty vector transfected GFP-infected condition. Data are means ± SEM (n = 4 independent hepatocyte cultures, P < 0.05 *: vs empty vector, #: vs PGC1α, two-tailed, unpaired Student’s t test). e Relative mRNA expression levels of indicated genes in primary hepatocytes 36 h post-transduction with indicated adenoviral vectors. Data are means ± SEM (representative experiment of n = 3 independent cultures, P < 0.05 *: vs GFP, #: vs PGC1α, 2-tailed, unpaired Student’s t test). f Relative transcript levels of mitochondria biogenesis factors and PPARα target Cpt1 in primary Vps15f/f hepatocytes in which Vps15 was depleted by expressing Cre recombinase using adenoviral vectors. Thirty-six hours after initial infection PGC1α was expressed for additional 24 h using adenoviral vectors. Data are means ± SEM (representative experiment of n = 3 independent hepatocyte cultures, P < 0.05 *: vs GFP/GFP, #: vs Cre/GFP, two-tailed, unpaired Student’s t test). g The OCR measured by SeaHorse Bioanalyzer in primary Vps15f/f hepatocytes in which Vps15 was depleted by expressing Cre recombinase using adenoviral vectors. Thirty-six hours after initial infection, cells were transduced with adenoviral vectors to express PGC1α or Vps15 for additional 24 h. First, OCR under basal conditions and then in response to sequential treatments is shown. Representative experiment of three independent hepatocyte cultures is presented. Dashed lines indicate the time of the addition of each reagent. Quantification of basal respiration, ATP production and maximal respiratory capacity are shown on the graphs (right panel). Data are means ± SEM (P < 0.05 *: vs GFP-infected cells, #: vs Cre-infected cells, two-tailed, unpaired Student’s t test)
