Journal: Physiological Reports

Article Title: Reduced mitochondrial mass and function add to age‐related susceptibility toward diet‐induced fatty liver in C57BL/6J mice

doi: 10.14814/phy2.12988

Figure Lengend Snippet: (A) Mitochondrial ROS production rates, monitored as H 2 O 2 released by liver mitochondria, isolated from adolescent (A), young adult (YA), and middle‐aged (MA) mice fed either a high fat or control diet for 9 weeks. Mitochondrial respiration was initiated with succinate/rotenone in absence of ADP. (B) Scatter plot analysis of mitochondrial ROS production rates plotted versus membrane potential or (D) oxygen consumption rate during leak respiration. Based on the regression equations shown in (B) and D), the respective residual ROS production rates were calculated in (C) and (E). n = 4–6 per age and diet group. Two‐Way ANOVA statistics are indicated with ** P < 0.01; *** P < 0.001; results of post hoc testing are indicated by # Significantly different to age‐matched control group; $ Significantly different to A within feeding group. Data are represented as means ± SD.

Article Snippet: Oxygen consumption of mitochondria (15 μg/mL) diluted in respiration buffer (120 mmol/L KCl, 5 mmol/L KH 2 PO 4 , 3 mmol/L HEPES, 1 mmol/L EGTA, all Carl Roth GmbH, with freshly added 0.3% fatty acid free BSA from Sigma Aldrich; pH 7.2) with succinate/rotenone (10 mmol/L/8 μmol/L) and subsequent injection of ADP (6 mmol/L) was recorded using the Seahorse XF 96 Analyzer (Agilent Technologies, Santa Clara, CA).

Techniques: Isolation, Control, Membrane

Journal: iScience

Article Title: Accumulated mtDNA mutations are linked to specific impairments in NADH-linked respiration

doi: 10.1016/j.isci.2026.115184

Figure Lengend Snippet: PolG Mut mitochondria possess selective impairments to NAD-linked Respiration (A) Schematic depiction of the substrates and inhibitors added during the OxPhos kinetics assay. Mitochondrial oxygen consumption ( J O 2 ) across OxPhos kinetics assay in mitochondria isolated from (B) BAT, (C) brain, (D) colon, (E) heart, (F) kidney, (G) Liver, (H) and lung tissue. Ratio of maximal complex I (CI) versus CII-supported mitochondrial J O 2 (I). N = 4–6 per group. Data are presented as mean ± SEM and analyzed using multiple unpaired t tests ( B–5H) or unpaired t test ( I), ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001. Substrates utilized are indicated as follows: creatine kinase (CK; 20 U/mL), ATP (5 mM), phospho-creatine (PCr; 1 mM), cytochrome c (Cyt C; 10 μM; pyruvate (Pyr; 5 mM), malate (Mal; 1 mM), octanoyl-carnitine (Oct; 0.2 mM), glutamate (Glut; 5 mM), rotenone (Rot; 0.5 μM), succinate (Succ; 5 mM) oligomycin (Oligo; 0.02 μM), malonate (Malo; 20 mM), calcium chloride (CaCl 2 ; 0.6 mM) glycerol-3-phosphate (G3P; 10 mM), and antimycin A (Ant A; 0.5 μM). Graphics were generated using BioRender.

Article Snippet: Following freeze fracture, 20μg of mitochondria were added to Respiration Buffer in the Oroboros O2k system followed by Cyt C (10μM).

Techniques: Isolation, Generated

Journal: iScience

Article Title: Accumulated mtDNA mutations are linked to specific impairments in NADH-linked respiration

doi: 10.1016/j.isci.2026.115184

Figure Lengend Snippet: Respiratory capacity within the electron transport system remains intact in PolG Mut mice (A–H) Schematic representing the maximal respiratory capacity protocol. Real-time oxygen consumption ( J O 2 ) in mitochondria isolated from (B) BAT, (C) brain, (D) colon, (E) heart, (F) kidney, (G) Liver, (H) and lung following serial titration of the mitochondrial uncoupling agent, carbonyl cyanide- p -trifluoromethoxyphenylhydrazone (FCCP, FC; 0.25 μM). (I) Maximal mitochondrial respiration achieved during FCCP titration. N = 5–6 per group, Data are presented as mean ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗∗ p < 0.0001 depict significant post hoc LSD by two-way ANOVA; main effect PolG ( B–4H) or unpaired t test ( I). Substrates utilized are indicated as follows: pyruvate (Pyr; 5 mM), malate (Mal; 1 mM), octanoyl-carnitine (Oct; 0.2 mM), succinate (Succ; 5 mM) cytochrome c (Cyt C, 10 μM). Graphics were generated using BioRender.

Article Snippet: Following freeze fracture, 20μg of mitochondria were added to Respiration Buffer in the Oroboros O2k system followed by Cyt C (10μM).

Techniques: Isolation, Titration, Generated

Journal: iScience

Article Title: Accumulated mtDNA mutations are linked to specific impairments in NADH-linked respiration

doi: 10.1016/j.isci.2026.115184

Figure Lengend Snippet: Mitochondrial respiratory phenotypes are maintained in permeabilized tissue (A) Tissue slices were permeabilized in saponin before assessment of oxygen consumption ( J O 2 ). (B) Intact colon, (C) heart, and (D) liver J O 2 under multiple substrate conditions. (E) Ratio of complex I (CI) versus CII-supported respiration. (F) Dose-response to FCCP titration and (G) maximal respiratory capacity in intact bone marrow-derived mononuclear cells (BMMCs). (H) Mitochondrial J O 2 during OxPhos kinetics technique in permeabilized BMMCs. (I) Ratio of CI versus CII-supported respiration in permeabilized BMMCs. N = 5 per group. Data are presented as mean ± SEM, p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001,∗∗∗∗ p < 0.00001 depict significant multiple unpaired t tests ( A–6C and 6G), and unpaired t test ( D–6F and 6H). Substrates utilized are indicated as follows: creatine kinase (CK; 20 U/mL), ATP (5 mM), phospho-creatine (PCr; 1 mM), cytochrome c (Cyt C; 10 μM), pyruvate (Pyr; 5 mM), malate (Mal; 1 mM), octanoyl-carnitine (Oct; 0.2 mM), glutamate (Glut; 5 mM), rotenone (Rot; 0.5 μM), succinate (Succ; 5 mM) oligomycin (Oligo; 0.02 μM), malonate (Malo; 20 mM), calcium chloride (CaCl 2 ; 0.6 mM) glycerol-3-phosphate (G3P; 10 mM) antimycin A (Ant A; 0.5 μM), and carbonyl cyanide- p -trifluoromethoxyphenylhydrazone (FCCP, FC; 0.25 μM). Graphics were generated using BioRender.

Article Snippet: Following freeze fracture, 20μg of mitochondria were added to Respiration Buffer in the Oroboros O2k system followed by Cyt C (10μM).

Techniques: Titration, Derivative Assay, Generated