Fig. 7
NOB restored mitochondrial supercomplex formation affected by HF feeding. Aged mice were fed with regular diet (RD), high-fat (HF) diet, or HF diet with NOB (HF.NOB). a Mitochondrial OXPHOS supercomplexes (SCs) analyzed by BN-PAGE. Representative BN-PAGE of digitonin-solubilized mitochondria purified from skeletal muscle of RD, HF, and HF-NOB-treated mice is shown. b Upper panel: enlarged view of the box area from a. Lower panel: quantification of SC5 band intensities (ZT6, n = 4, 4, and 3 for RD, HF, and HF.NOB, respectively; ZT18, n = 5, 5, and 4 for RD, HF, and HF.NOB, respectively). c In-gel activity assay with Complex IV and subsequently with Complex I substrates performed as described in Methods. Brown and violet color bands indicate CIV and CI activity, respectively. d Western blot analysis of the respiratory Complexes I, III and IV isolated from skeletal muscle mitochondria in aged mice. Western blotting was performed as described in Methods. Lane 1: antibody to CI subunit 8, pseudo-colored blue; lane 2: antibody to CIII, subunit 2 (QCR2, Cor2), pseudo-colored red; lane 3: antibody to CIV subunit I (Cox1), pseudo-colored green; lane 4: overlay of lanes 2 and 3. Of note, in SC labels, n reflects the number of individual complexes in SCs, not the level of their oligomerization. *p < 0.05, one-way ANOVA. Data are presented as mean ± SEM in bar graphs
