Figure 4. The lipogenic switch alters the redox system and lipid‐related OXPHOS components.

Data on wild‐type (wt, black bars) and Low_OXPHOS (orange bars) mice are shown.

• A
  v‐WAT iTRAQ ratio of proteins from BCAA catabolism (wt, n = 4; Low_OXPHOS, n = 4). Higher intensities of blue colors represent lower Low_OXPHOS/wt expression ratios.
• B
  Odd chain FAs in Skm and WAT from wt (n = 8) or Low_OXPHOS (n = 8) mice.
• C, D
  Representative WB expression (C) and iTRAQ analysis (D) of v‐WAT proteins from lipid metabolism. Each sample contains extracts from 3 mice. The histogram shows the quantifications (wt, n = 6; Low_OXPHOS, n = 6).
• E
  v‐WAT weight expressed as a percentage of the wt (wt, n = 12; Low_OXPHOS, n = 12).
• F
  Quantitative lipidomics in v‐WAT. The color scale (brown to blue) in the heat map represents Low_OXPHOS/wt amounts of specific lipid species. A P‐value ≤ 0.05 was considered statistically significant (red). Each sample is a pool from 4 mouse extracts, n = 8/genotype.
• G
  v‐WAT amounts of triacylglycerides (TAGs) and diacylglycerides (DAGs) (wt, n = 8; Low_OXPHOS, n = 8).
• H
  Higher saturated lipid species in Low_OXPHOS mice compared to wt (wt, n = 8; Low_OXPHOS, n = 8).
• I
  MitoSox staining in myocytes expressing or not ATPIF1_H49K. The right histogram shows the quantification of mitochondrial ROS. Bars are the mean ± SEM of n = 3 experiments, 12 replicas/condition.
• J
  Representative WB of Skm lipid peroxidation and redox system proteins. Two samples per condition are shown. Each sample contains extracts from 3 mice. Histograms represent quantification (wt, n = 6; Low_OXPHOS, n = 6). 4‐Hydroxynonenal (4HN), peroxiredoxin 2, 3 and 6 (PRX), superoxide dismutase 1 and 2 (SOD), catalase (CATA) and glutathione reductase (GSR) immunoblots are shown. βF1 is presented as a loading control.
• K
  Representative WB of Skm OXPHOS and related system proteins. Two samples per condition are shown. Each sample contains extracts from 3 mice. Histograms represent quantification (wt, n = 6; Low_OXPHOS, n = 6). NDUFA9 (CI), SDHA and B (CII), ETF subunits A and B, ETFDH, CoreII (CIII), subunit 1 (CIV), βF1(CV) and hATPIF1 immunoblots are shown. Tubulin is presented as a loading control. See also Fig EV5.
• L
  Skm iTRAQ ratio of FADH₂‐binding proteins (wt, n = 12; Low_OXPHOS, n = 12). A higher intensity of red color represents a higher Low_OXPHOS/wt expression ratio.
• M
  Oxidized FAD levels and the reduced/oxidized FAD ratio in Skm extracts (wt, n = 8; Low_OXPHOS n = 8).

Data information: Bars are the mean ± SEM of the indicated (n) mice/genotype. *P < 0.05 when compared to wt by Student's t‐test. See also Figs EV4 and EV5, and Table EV3.Source data are available online for this figure.