citrate synthase Search Results


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  • 99
    Millipore citrate synthase activity assay
    Effect of suppressing NFkB activation in response to cellular fuel overloading on mitochondrial morphology, mitochondrial proteins and gene expression. L6 myotubes were incubated with GLC (5 mM), PA (0.4 mM), 2DG (5 mM) and BI605906 (10 μM) for 16 h in the combinations indicated in the various experimental data panels prior to a analysis and quantification of mitochondrial morphology using Mitotracker green (Mitospy) by confocal microscopy (the scale bar represents 5 μm), b mitochondrial DNA copy number by qPCR, c citrate <t>synthase</t> (CS) activity and ( d , e ), analysis of mitochondria protein and mRNA abundance (UCP3, ANT1, PGC1α, SDHA, and COX4.1) which was normalised to GAPDH. All graphical bar data are presented as mean ± SEM from four separate experiments. Asterisks indicate a significant change ( P
    Citrate Synthase Activity Assay, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 61 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore citrate synthase
    Chaperone assay of GhHSP proteins. ( A ) Effect of control protein BSA and inhibitor Nov on activity of Citrate <t>synthase</t> (CS) ( B ) Effect of control protein BSA and Pif on activity of CS. ( C ) Effect of Human HSP90 on activity of CS, with or without Nov. ( D ) Effect of Human HSP70 on activity of CS, with or without Pif. ( E ) Effect of GhHSP90 on activity of CS, with or without Nov. ( F ) Effect of GhHSP70 on activity of CS, with or without Pif. ( G ) The mean CS activity at the end point of A, C and E relative to CS-thermosprotection in the presence of HSP90 protein. ( H ) The mean CS activity at the end point of B, D and F relative to CS-thermosprotection in the presence of HSP70 protein. The asterisks represent statistical significance between two independent experiments (*p-value ≤ 0.05, **p-value ≤ 0.01).
    Citrate Synthase, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 449 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore citrate synthase assay kit
    Total pool of phosphocreatine, creatine, PCr + Cr, ATP, total adenine nucleotides (TAN) ( A ) and citrate <t>synthase</t> activity ( B ) in rat hearts. Data are expressed as means ± SEM and one-way ANOVA was used to compare the groups. * p
    Citrate Synthase Assay Kit, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 665 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Abcam citrate synthase
    OPA 1 downregulation decreases mitochondrial respiration, induces the nuclear translocation of NRF 2, and increases both catalase quantity and activity in cortical neurons ex vivo. (A) Representative immunoblots and histograms showing protein levels of OPA 1 (inner membrane), citrate <t>synthase</t> (matrix), HSP 60 (matrix), VDAC (outer membrane), and TOM 20 (outer membrane) relative to actin in si OPA 1‐ (gray bars) and siCtrl‐transfected (white bars) neurons. Only OPA 1 protein quantity is drastically decreased (0.23 ± 0.06 AU) in si OPA 1 neurons when compared to controls (1.08 ± 0.16 AU). Results are expressed as mean ± SEM ( n = 5–8). Statistical significance was determined by Student's paired t ‐test, *** P
    Citrate Synthase, supplied by Abcam, used in various techniques. Bioz Stars score: 95/100, based on 229 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Cayman Chemical citrate synthase activity
    Loss of Chrna2 reduces the adaptive thermogenic capacity of inguinal adipose tissue. ( a ) Changes in body weight of wild type (WT) and Chrna2 KO mice following 2 weeks cold exposure (CE) at 10°C (n = 22 for WT, 25 for KO). ( b ) Inguinal (IWAT), visceral (VWAT), and interscapular brown (BAT) fat mass in WT and Chrna2 KO mice after CE (n = 22 for WT, 31 for KO). ( c, d ) qPCR analyses of thermogenic genes ( c ) and mitochondrial genes ( d ) in IWAT from WT and Chrna2 KO mice after CE (n = 16 for WT, 15 for KO). ( e ) mitochondrial DNA content in IWAT of WT and Chrna2 KO mice following CE (n = 8 per group). ( f ) Citrate <t>synthase</t> activity in IWAT of cold exposed WT and Chrna2 KO mice (n = 10 per group). ( g ) Immunoblot analyses of UCP1, the mitochondrial marker COXIV and OxPhos components in IWAT of WT and Chrna2 KO mice after CE. Representative images are shown. ( h ) Oxygen consumption rate (OCR) in freshly isolated inguinal fat tissue from cold exposed WT and Chrna2 KO mice in the absence (basal) or presence of oligomycin (n = 6 for WT, 7 for KO). ( i ) qPCR analyses of thermogenic markers in the interscapular BAT from WT and Chrna2 KO mice following 2 weeks CE (n = 16 per group). Data are presented as mean ± s.e.m. Data were analyzed by a two-tailed Student’s t -test ( a–f,h,i ). * P
    Citrate Synthase Activity, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 90/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Alpha Diagnostics citrate synthase
    Severe iron deficiency, hexokinase and citrate <t>synthase</t> expression in gastrocnemius muscle. A . Hexokinase expression increased significantly with iron deficiency (ǂ indicates statistical significance, p
    Citrate Synthase, supplied by Alpha Diagnostics, used in various techniques. Bioz Stars score: 92/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology citrate synthase
    Effect of extended aging on citrate <t>synthase</t> and peroxiredoxin III (Prx III) amounts in rat liver. Extended aging did not induce any significant change in citrate synthase and Prx III amounts. A . Representative western blot carried out in four rats from each assayed group. The bands from top to bottom show, respectively, the signals from citrate synthase, Prx III and VDAC. B. The histogram shows the relative amounts of citrate synthase and Prx III in AL-32 rats, determined by densitometry analysis of the results from triplicated western blots experiments, compared to AL-28 rats. The densitometric value of OD units of every citrate synthase and Prx III band was related to the number of OD units of the respective band of VDAC for each analyzed sample. Bars represent the mean ((±SE) of the relative (citrate synthase/VDAC, Prx III/VDAC) values obtained from each AL-28 and AL-32 rat. Comparisons were made with respects to the value of the AL-28 rats, fixed as 1. n = number of analyzed animals.
    Citrate Synthase, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Abcam citrate synthase activity assay kit
    Atgl controls mitochondrial functionality in muscle cells. (A) Western blot analysis of Atgl in total homogenates of GSCN of mice fed with normal diet (ND) or high fat diet (HFD). Immunoblots reported are representative of three mice per group out of five giving similar results. Below are reported the densitometric analyses of the immunoreactive bands normalized to Ponceau staining (entire line). (B) mRNA expression analysis in gastrocnemius (GSCN) upon treatment with HFD. (C) Western blot analysis of Atgl in C2C12 myotubes transfected with a siRNA against Atgl (Atgl-) or with a scramble siRNA (Scr) ( left panel ). Immunoblots reported are representative of one experiment out of three giving similar results. Tubulin was used as loading control. Measurement of mitochondrial membrane potential (ΔΨM) and citrate <t>synthase</t> activity in crude mitochondrial fractions of Atgl- C2C12 myotubes ( right panel ). (D) Lipid content measured by Oil Red O staining ( right panel ) in C2C12 myotubes transfected with the Atgl cDNA (Atgl+) or with empty vector (Empty) and treated with 300 μM palmitic acid (PA) for 72h. Atgl immunoblot ( left panel ) is representative of three independent experiments giving similar results. Tubulin was used as loading control. (E) Measurement of mitochondrial membrane potential (ΔΨM) in crude mitochondrial fractions of Atgl+ C2C12 myotubes. (F) Measurement of citrate synthase activity in crude mitochondrial fractions of Atgl+ C2C12 myotubes. All data are expressed as mean ±S.D. (n = 5 mice/group). In vitro data are representative of at least three independent experiments. Student’s t-test was used for two groups comparisons (A-C). One-way ANOVA analysis followed by Turkey’s test corrections was used for multiple comparisons (D-F).
    Citrate Synthase Activity Assay Kit, supplied by Abcam, used in various techniques. Bioz Stars score: 96/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    BioVision citrate synthase activity colorimetric assay kit
    The effects of exercise on body weight at weeks 1–8 ( n = 20 animals per group) (A), intermittent hypoxia on body weight at weeks 9–10 ( n = 5 animals per group) (B) and exercise on citrate <t>synthase</t> activity ( n = 5 animals per group) (C) . # p
    Citrate Synthase Activity Colorimetric Assay Kit, supplied by BioVision, used in various techniques. Bioz Stars score: 96/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam anti citrate synthetase antibody
    The effects of exercise on body weight at weeks 1–8 ( n = 20 animals per group) (A), intermittent hypoxia on body weight at weeks 9–10 ( n = 5 animals per group) (B) and exercise on citrate <t>synthase</t> activity ( n = 5 animals per group) (C) . # p
    Anti Citrate Synthetase Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Cell Signaling Technology Inc citrate synthase
    The effects of exercise on body weight at weeks 1–8 ( n = 20 animals per group) (A), intermittent hypoxia on body weight at weeks 9–10 ( n = 5 animals per group) (B) and exercise on citrate <t>synthase</t> activity ( n = 5 animals per group) (C) . # p
    Citrate Synthase, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cayman Chemical mitocheck citrate synthase activity assay kit
    The effects of exercise on body weight at weeks 1–8 ( n = 20 animals per group) (A), intermittent hypoxia on body weight at weeks 9–10 ( n = 5 animals per group) (B) and exercise on citrate <t>synthase</t> activity ( n = 5 animals per group) (C) . # p
    Mitocheck Citrate Synthase Activity Assay Kit, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 99/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mitocheck citrate synthase activity assay kit/product/Cayman Chemical
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    98
    Cayman Chemical citrate synthase cs activity
    mtCaMKII activates TCA cycle dehydrogenases and augments resting NADH. a Pyruvate dehydrogenase (PDH) activity in isolated mitochondria from WT ( n = 7) and mtCaMKII ( n = 9) hearts. b TCA cycle enzyme activities in permeabilized mitochondria from WT ( n = 7) and mtCaMKII ( n = 9) hearts, CS: citrate <t>synthase,</t> IDH: isocitrate dehydrogenase, α-KGDH: α-ketoglutarate dehydrogenase, SL: succinate-CoA ligase, SDH: succinate dehydrogenase, MDH: malate dehydrogenase. c Representative NADH imaging of isolated ventricular myocytes before pacing (calibration bar = 50 μm). d Quantification of NADH autofluorescence in isolated ventricular myocytes before and after pacing from WT ( n = 29 myocytes from two hearts) and mtCaMKII ( n = 52 myocytes from three hearts) hearts. Maximum (100%) NADH was measured in the presence of NaCN, and minimum (0%) in the presence of FCCP. Cells from the same genotype were paired in comparisons of before and after pacing. e Quantification of NAD + and NADH in hearts from WT ( n = 4) and mtCaMKII ( n = 4) mice. f Western blot and g summary data for acetylated proteins normalized to VDAC1 in isolated mitochondria from WT ( n = 4) and mtCaMKII ( n = 4) hearts. Data are represented as mean ± SEM, significance was determined using a two-tailed Student’s t test. **** P
    Citrate Synthase Cs Activity, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 98/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    GeneTex citrate synthase
    Influence of SIRT3 depletion on mitochondrial activity. A) Respiration rates in C2C12- LucshRNA (shCTL) and C2C12-SIRT3shRNA (shSIRT3) at cell proliferation (P), cell confluence (C) and 3 days of differentiation (D3). Values are normalized relatively to total protein levels. B) Citrate <t>synthase,</t> complex II and cytochrome c oxidase maximal activities in shCTL and shSIRT3 cells at cell proliferation (P), cell confluence (C) and 3 days of differentiation (D3). Results are expressed as mean ± SD from five independent experiments. ANOVA main effect: #P
    Citrate Synthase, supplied by GeneTex, used in various techniques. Bioz Stars score: 92/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effect of suppressing NFkB activation in response to cellular fuel overloading on mitochondrial morphology, mitochondrial proteins and gene expression. L6 myotubes were incubated with GLC (5 mM), PA (0.4 mM), 2DG (5 mM) and BI605906 (10 μM) for 16 h in the combinations indicated in the various experimental data panels prior to a analysis and quantification of mitochondrial morphology using Mitotracker green (Mitospy) by confocal microscopy (the scale bar represents 5 μm), b mitochondrial DNA copy number by qPCR, c citrate synthase (CS) activity and ( d , e ), analysis of mitochondria protein and mRNA abundance (UCP3, ANT1, PGC1α, SDHA, and COX4.1) which was normalised to GAPDH. All graphical bar data are presented as mean ± SEM from four separate experiments. Asterisks indicate a significant change ( P

    Journal: Cellular and Molecular Life Sciences

    Article Title: Proinflammatory NFkB signalling promotes mitochondrial dysfunction in skeletal muscle in response to cellular fuel overloading

    doi: 10.1007/s00018-019-03148-8

    Figure Lengend Snippet: Effect of suppressing NFkB activation in response to cellular fuel overloading on mitochondrial morphology, mitochondrial proteins and gene expression. L6 myotubes were incubated with GLC (5 mM), PA (0.4 mM), 2DG (5 mM) and BI605906 (10 μM) for 16 h in the combinations indicated in the various experimental data panels prior to a analysis and quantification of mitochondrial morphology using Mitotracker green (Mitospy) by confocal microscopy (the scale bar represents 5 μm), b mitochondrial DNA copy number by qPCR, c citrate synthase (CS) activity and ( d , e ), analysis of mitochondria protein and mRNA abundance (UCP3, ANT1, PGC1α, SDHA, and COX4.1) which was normalised to GAPDH. All graphical bar data are presented as mean ± SEM from four separate experiments. Asterisks indicate a significant change ( P

    Article Snippet: Citrate synthase (CS) activity was measured using a kit purchased from Sigma-Aldrich/UK (MAK193).

    Techniques: Activation Assay, Expressing, Incubation, Gas Chromatography, Confocal Microscopy, Real-time Polymerase Chain Reaction, Activity Assay

    Chaperone assay of GhHSP proteins. ( A ) Effect of control protein BSA and inhibitor Nov on activity of Citrate synthase (CS) ( B ) Effect of control protein BSA and Pif on activity of CS. ( C ) Effect of Human HSP90 on activity of CS, with or without Nov. ( D ) Effect of Human HSP70 on activity of CS, with or without Pif. ( E ) Effect of GhHSP90 on activity of CS, with or without Nov. ( F ) Effect of GhHSP70 on activity of CS, with or without Pif. ( G ) The mean CS activity at the end point of A, C and E relative to CS-thermosprotection in the presence of HSP90 protein. ( H ) The mean CS activity at the end point of B, D and F relative to CS-thermosprotection in the presence of HSP70 protein. The asterisks represent statistical significance between two independent experiments (*p-value ≤ 0.05, **p-value ≤ 0.01).

    Journal: Scientific Reports

    Article Title: Inhibition of Heat Shock proteins HSP90 and HSP70 induce oxidative stress, suppressing cotton fiber development

    doi: 10.1038/s41598-018-21866-0

    Figure Lengend Snippet: Chaperone assay of GhHSP proteins. ( A ) Effect of control protein BSA and inhibitor Nov on activity of Citrate synthase (CS) ( B ) Effect of control protein BSA and Pif on activity of CS. ( C ) Effect of Human HSP90 on activity of CS, with or without Nov. ( D ) Effect of Human HSP70 on activity of CS, with or without Pif. ( E ) Effect of GhHSP90 on activity of CS, with or without Nov. ( F ) Effect of GhHSP70 on activity of CS, with or without Pif. ( G ) The mean CS activity at the end point of A, C and E relative to CS-thermosprotection in the presence of HSP90 protein. ( H ) The mean CS activity at the end point of B, D and F relative to CS-thermosprotection in the presence of HSP70 protein. The asterisks represent statistical significance between two independent experiments (*p-value ≤ 0.05, **p-value ≤ 0.01).

    Article Snippet: The aggregation of 0.5 µM citrate synthase (from Porcine heart; Sigma) was induced at 43 °C in 50 mM HEPES (pH 8.0), with or without 1.8 µM proteins (BSA or HSP90/70 (Human, Sigma) or GhHSP90/70) and with or without HSP90/70 inhibitors Nov or Pif respectively.

    Techniques: Activity Assay

    (A) Changes in the near-UV CD signal of BiP at 266nm as a function of varying ATP concentration. (B) Changes in ATPase activity of protein before and after chemical modification. Blue bar is ATP only. (C) Changes in fluorescence anisotropy of peptide HTFPAVL at 482nm as a function of varying BiP concentration. (D) Changes in light scattering at 350 nm due to the aggregation of citrate synthase as a function of incubation time at 43°C. Blue indicates citrate synthase alone. In all panels, black and red colors indicate the data for wild-type BiP and chemically modified BiP, respectively.

    Journal: PLoS ONE

    Article Title: Biophysical analysis of the effect of chemical modification by 4-oxononenal on the structure, stability, and function of binding immunoglobulin protein (BiP)

    doi: 10.1371/journal.pone.0183975

    Figure Lengend Snippet: (A) Changes in the near-UV CD signal of BiP at 266nm as a function of varying ATP concentration. (B) Changes in ATPase activity of protein before and after chemical modification. Blue bar is ATP only. (C) Changes in fluorescence anisotropy of peptide HTFPAVL at 482nm as a function of varying BiP concentration. (D) Changes in light scattering at 350 nm due to the aggregation of citrate synthase as a function of incubation time at 43°C. Blue indicates citrate synthase alone. In all panels, black and red colors indicate the data for wild-type BiP and chemically modified BiP, respectively.

    Article Snippet: Porcine citrate synthase (Cat# C3260-1KU) and malachite green oxalate salt (Cat# M9015) were bought from Sigma Aldrich, St Louis, MO.

    Techniques: Concentration Assay, Activity Assay, Modification, Fluorescence, Incubation

    Total pool of phosphocreatine, creatine, PCr + Cr, ATP, total adenine nucleotides (TAN) ( A ) and citrate synthase activity ( B ) in rat hearts. Data are expressed as means ± SEM and one-way ANOVA was used to compare the groups. * p

    Journal: Nutrients

    Article Title: Protective Effect of Resveratrol against Ischemia-Reperfusion Injury via Enhanced High Energy Compounds and eNOS-SIRT1 Expression in Type 2 Diabetic Female Rat Heart

    doi: 10.3390/nu11010105

    Figure Lengend Snippet: Total pool of phosphocreatine, creatine, PCr + Cr, ATP, total adenine nucleotides (TAN) ( A ) and citrate synthase activity ( B ) in rat hearts. Data are expressed as means ± SEM and one-way ANOVA was used to compare the groups. * p

    Article Snippet: Citrate synthase activity was evaluated using the citrate synthase assay kit (CS0720, Sigma-Aldrich, St. Louis, MO, USA).

    Techniques: Polymerase Chain Reaction, Activity Assay

    Citrate synthase activity normalized to total protein for CON, BFRRE, and HLRE groups. Data are presented as means ± SD. Overall effects are designated in the upper left corner of the graph.

    Journal: Frontiers in Physiology

    Article Title: Skeletal Muscle Mitochondrial Protein Synthesis and Respiration Increase With Low-Load Blood Flow Restricted as Well as High-Load Resistance Training

    doi: 10.3389/fphys.2018.01796

    Figure Lengend Snippet: Citrate synthase activity normalized to total protein for CON, BFRRE, and HLRE groups. Data are presented as means ± SD. Overall effects are designated in the upper left corner of the graph.

    Article Snippet: Citrate synthase activity was measured spectrophotometrically using a citrate synthase activity assay kit (Cat. CS0720, Sigma-Aldrich, St. Louis, MO, United States).

    Techniques: Activity Assay

    OPA 1 downregulation decreases mitochondrial respiration, induces the nuclear translocation of NRF 2, and increases both catalase quantity and activity in cortical neurons ex vivo. (A) Representative immunoblots and histograms showing protein levels of OPA 1 (inner membrane), citrate synthase (matrix), HSP 60 (matrix), VDAC (outer membrane), and TOM 20 (outer membrane) relative to actin in si OPA 1‐ (gray bars) and siCtrl‐transfected (white bars) neurons. Only OPA 1 protein quantity is drastically decreased (0.23 ± 0.06 AU) in si OPA 1 neurons when compared to controls (1.08 ± 0.16 AU). Results are expressed as mean ± SEM ( n = 5–8). Statistical significance was determined by Student's paired t ‐test, *** P

    Journal: Annals of Clinical and Translational Neurology

    Article Title: Loss of functional OPA1 unbalances redox state: implications in dominant optic atrophy pathogenesis

    doi: 10.1002/acn3.305

    Figure Lengend Snippet: OPA 1 downregulation decreases mitochondrial respiration, induces the nuclear translocation of NRF 2, and increases both catalase quantity and activity in cortical neurons ex vivo. (A) Representative immunoblots and histograms showing protein levels of OPA 1 (inner membrane), citrate synthase (matrix), HSP 60 (matrix), VDAC (outer membrane), and TOM 20 (outer membrane) relative to actin in si OPA 1‐ (gray bars) and siCtrl‐transfected (white bars) neurons. Only OPA 1 protein quantity is drastically decreased (0.23 ± 0.06 AU) in si OPA 1 neurons when compared to controls (1.08 ± 0.16 AU). Results are expressed as mean ± SEM ( n = 5–8). Statistical significance was determined by Student's paired t ‐test, *** P

    Article Snippet: The membranes were probed with various primary antibodies: anti‐OPA1 (1/300, BD Bioscience, USA), anti‐actin (1/25000, Chemicon, Merck Millipore, UK), anti‐HSP60 (1/8000, Sigma‐Aldrich, UK), anti‐citrate synthase (1/3000, Abcam, Cambridge, USA), anti‐VDAC (1/1000, Abcam, Cambridge, USA), anti‐TOM20 (1/2000, Abcam, Cambridge, USA), anti‐aconitase (1/500, Abcam, Cambridge, USA), anti‐SOD1 and anti‐SOD2 (1/2000, Epitomics, Abcam, Cambridge, USA), anti‐catalase (1/3000, Abcam, Cambridge, USA), anti‐NQO1 (1/3000, Abcam, Cambridge, USA), anti‐mitofilin (1/1000 Abcam, Cambridge, USA), anti‐MFN1 (1/1000 , Abnova, Taipei City, Taiwan), and anti‐GSTP1 (1/8000, Oxford Biochemical Research, Euromedex, France), and incubated overnight at 4°C in blocking buffer.

    Techniques: Translocation Assay, Activity Assay, Ex Vivo, Western Blot, Transfection

    Loss of Chrna2 reduces the adaptive thermogenic capacity of inguinal adipose tissue. ( a ) Changes in body weight of wild type (WT) and Chrna2 KO mice following 2 weeks cold exposure (CE) at 10°C (n = 22 for WT, 25 for KO). ( b ) Inguinal (IWAT), visceral (VWAT), and interscapular brown (BAT) fat mass in WT and Chrna2 KO mice after CE (n = 22 for WT, 31 for KO). ( c, d ) qPCR analyses of thermogenic genes ( c ) and mitochondrial genes ( d ) in IWAT from WT and Chrna2 KO mice after CE (n = 16 for WT, 15 for KO). ( e ) mitochondrial DNA content in IWAT of WT and Chrna2 KO mice following CE (n = 8 per group). ( f ) Citrate synthase activity in IWAT of cold exposed WT and Chrna2 KO mice (n = 10 per group). ( g ) Immunoblot analyses of UCP1, the mitochondrial marker COXIV and OxPhos components in IWAT of WT and Chrna2 KO mice after CE. Representative images are shown. ( h ) Oxygen consumption rate (OCR) in freshly isolated inguinal fat tissue from cold exposed WT and Chrna2 KO mice in the absence (basal) or presence of oligomycin (n = 6 for WT, 7 for KO). ( i ) qPCR analyses of thermogenic markers in the interscapular BAT from WT and Chrna2 KO mice following 2 weeks CE (n = 16 per group). Data are presented as mean ± s.e.m. Data were analyzed by a two-tailed Student’s t -test ( a–f,h,i ). * P

    Journal: Nature medicine

    Article Title: An immune-beige adipocyte communication via nicotinic acetylcholine receptor signaling

    doi: 10.1038/s41591-018-0032-8

    Figure Lengend Snippet: Loss of Chrna2 reduces the adaptive thermogenic capacity of inguinal adipose tissue. ( a ) Changes in body weight of wild type (WT) and Chrna2 KO mice following 2 weeks cold exposure (CE) at 10°C (n = 22 for WT, 25 for KO). ( b ) Inguinal (IWAT), visceral (VWAT), and interscapular brown (BAT) fat mass in WT and Chrna2 KO mice after CE (n = 22 for WT, 31 for KO). ( c, d ) qPCR analyses of thermogenic genes ( c ) and mitochondrial genes ( d ) in IWAT from WT and Chrna2 KO mice after CE (n = 16 for WT, 15 for KO). ( e ) mitochondrial DNA content in IWAT of WT and Chrna2 KO mice following CE (n = 8 per group). ( f ) Citrate synthase activity in IWAT of cold exposed WT and Chrna2 KO mice (n = 10 per group). ( g ) Immunoblot analyses of UCP1, the mitochondrial marker COXIV and OxPhos components in IWAT of WT and Chrna2 KO mice after CE. Representative images are shown. ( h ) Oxygen consumption rate (OCR) in freshly isolated inguinal fat tissue from cold exposed WT and Chrna2 KO mice in the absence (basal) or presence of oligomycin (n = 6 for WT, 7 for KO). ( i ) qPCR analyses of thermogenic markers in the interscapular BAT from WT and Chrna2 KO mice following 2 weeks CE (n = 16 per group). Data are presented as mean ± s.e.m. Data were analyzed by a two-tailed Student’s t -test ( a–f,h,i ). * P

    Article Snippet: Citrate synthase activity in the supernatant was measured with a citrate synthase activity assay kit according to manufacturer’s instructions (Cayman chemical).

    Techniques: Mouse Assay, Real-time Polymerase Chain Reaction, Activity Assay, Marker, Isolation, Two Tailed Test

    Severe iron deficiency, hexokinase and citrate synthase expression in gastrocnemius muscle. A . Hexokinase expression increased significantly with iron deficiency (ǂ indicates statistical significance, p

    Journal: Nutrition & Metabolism

    Article Title: Iron deficiency causes a shift in AMP-activated protein kinase (AMPK) subunit composition in rat skeletal muscle

    doi: 10.1186/1743-7075-9-104

    Figure Lengend Snippet: Severe iron deficiency, hexokinase and citrate synthase expression in gastrocnemius muscle. A . Hexokinase expression increased significantly with iron deficiency (ǂ indicates statistical significance, p

    Article Snippet: Western blot analysis was also performed on mixed quadriceps muscle for citrate synthase (1:10,000 from Alpha Diagnostic; cat. no. CISY11-A).

    Techniques: Expressing

    Effect of extended aging on citrate synthase and peroxiredoxin III (Prx III) amounts in rat liver. Extended aging did not induce any significant change in citrate synthase and Prx III amounts. A . Representative western blot carried out in four rats from each assayed group. The bands from top to bottom show, respectively, the signals from citrate synthase, Prx III and VDAC. B. The histogram shows the relative amounts of citrate synthase and Prx III in AL-32 rats, determined by densitometry analysis of the results from triplicated western blots experiments, compared to AL-28 rats. The densitometric value of OD units of every citrate synthase and Prx III band was related to the number of OD units of the respective band of VDAC for each analyzed sample. Bars represent the mean ((±SE) of the relative (citrate synthase/VDAC, Prx III/VDAC) values obtained from each AL-28 and AL-32 rat. Comparisons were made with respects to the value of the AL-28 rats, fixed as 1. n = number of analyzed animals.

    Journal: Experimental gerontology

    Article Title: “What makes some rats live so long?” The mitochondrial contribution to longevity through balance of mitochondrial dynamics and mtDNA content

    doi: 10.1016/j.exger.2016.09.010

    Figure Lengend Snippet: Effect of extended aging on citrate synthase and peroxiredoxin III (Prx III) amounts in rat liver. Extended aging did not induce any significant change in citrate synthase and Prx III amounts. A . Representative western blot carried out in four rats from each assayed group. The bands from top to bottom show, respectively, the signals from citrate synthase, Prx III and VDAC. B. The histogram shows the relative amounts of citrate synthase and Prx III in AL-32 rats, determined by densitometry analysis of the results from triplicated western blots experiments, compared to AL-28 rats. The densitometric value of OD units of every citrate synthase and Prx III band was related to the number of OD units of the respective band of VDAC for each analyzed sample. Bars represent the mean ((±SE) of the relative (citrate synthase/VDAC, Prx III/VDAC) values obtained from each AL-28 and AL-32 rat. Comparisons were made with respects to the value of the AL-28 rats, fixed as 1. n = number of analyzed animals.

    Article Snippet: 10 μg of mitochondrial proteins were used for western blot analysis as described in ( ) with the following primary antibodies dilutions used: TFAM (1:50,000), VDAC (1:50,000, Abcam), OGG1 (1:2500, Abcam), APE1 (1:5000, Abcam), MFN2 (1:5000, Abnova), DRP1 (1:2500, Abnova), Cyt c (1:500, Pharmingen), Citrate Synthase (1:100,000, Santa Cruz), Lon Protease (1:10,000) and Prx III (1:50,000, AbFrontier).

    Techniques: Western Blot

    Atgl controls mitochondrial functionality in muscle cells. (A) Western blot analysis of Atgl in total homogenates of GSCN of mice fed with normal diet (ND) or high fat diet (HFD). Immunoblots reported are representative of three mice per group out of five giving similar results. Below are reported the densitometric analyses of the immunoreactive bands normalized to Ponceau staining (entire line). (B) mRNA expression analysis in gastrocnemius (GSCN) upon treatment with HFD. (C) Western blot analysis of Atgl in C2C12 myotubes transfected with a siRNA against Atgl (Atgl-) or with a scramble siRNA (Scr) ( left panel ). Immunoblots reported are representative of one experiment out of three giving similar results. Tubulin was used as loading control. Measurement of mitochondrial membrane potential (ΔΨM) and citrate synthase activity in crude mitochondrial fractions of Atgl- C2C12 myotubes ( right panel ). (D) Lipid content measured by Oil Red O staining ( right panel ) in C2C12 myotubes transfected with the Atgl cDNA (Atgl+) or with empty vector (Empty) and treated with 300 μM palmitic acid (PA) for 72h. Atgl immunoblot ( left panel ) is representative of three independent experiments giving similar results. Tubulin was used as loading control. (E) Measurement of mitochondrial membrane potential (ΔΨM) in crude mitochondrial fractions of Atgl+ C2C12 myotubes. (F) Measurement of citrate synthase activity in crude mitochondrial fractions of Atgl+ C2C12 myotubes. All data are expressed as mean ±S.D. (n = 5 mice/group). In vitro data are representative of at least three independent experiments. Student’s t-test was used for two groups comparisons (A-C). One-way ANOVA analysis followed by Turkey’s test corrections was used for multiple comparisons (D-F).

    Journal: PLoS ONE

    Article Title: Time-controlled fasting prevents aging-like mitochondrial changes induced by persistent dietary fat overload in skeletal muscle

    doi: 10.1371/journal.pone.0195912

    Figure Lengend Snippet: Atgl controls mitochondrial functionality in muscle cells. (A) Western blot analysis of Atgl in total homogenates of GSCN of mice fed with normal diet (ND) or high fat diet (HFD). Immunoblots reported are representative of three mice per group out of five giving similar results. Below are reported the densitometric analyses of the immunoreactive bands normalized to Ponceau staining (entire line). (B) mRNA expression analysis in gastrocnemius (GSCN) upon treatment with HFD. (C) Western blot analysis of Atgl in C2C12 myotubes transfected with a siRNA against Atgl (Atgl-) or with a scramble siRNA (Scr) ( left panel ). Immunoblots reported are representative of one experiment out of three giving similar results. Tubulin was used as loading control. Measurement of mitochondrial membrane potential (ΔΨM) and citrate synthase activity in crude mitochondrial fractions of Atgl- C2C12 myotubes ( right panel ). (D) Lipid content measured by Oil Red O staining ( right panel ) in C2C12 myotubes transfected with the Atgl cDNA (Atgl+) or with empty vector (Empty) and treated with 300 μM palmitic acid (PA) for 72h. Atgl immunoblot ( left panel ) is representative of three independent experiments giving similar results. Tubulin was used as loading control. (E) Measurement of mitochondrial membrane potential (ΔΨM) in crude mitochondrial fractions of Atgl+ C2C12 myotubes. (F) Measurement of citrate synthase activity in crude mitochondrial fractions of Atgl+ C2C12 myotubes. All data are expressed as mean ±S.D. (n = 5 mice/group). In vitro data are representative of at least three independent experiments. Student’s t-test was used for two groups comparisons (A-C). One-way ANOVA analysis followed by Turkey’s test corrections was used for multiple comparisons (D-F).

    Article Snippet: Citrate synthase activity was measured by using a Citrate Synthase Activity Assay Kit (Abcam, Cambridge, UK).

    Techniques: Western Blot, Mouse Assay, Staining, Expressing, Transfection, Activity Assay, Plasmid Preparation, In Vitro

    HFD induces mitochondrial damage and dysfunction in SkM. (A) Basal oxygen consumption measured by a polarographic method in gastrocnemius (GSCN) and tibialis anterior muscle (TA) of mice fed with normal diet (ND) or high fat diet (HFD). (B) Citrate synthase activity ( left panel ) and mitochondrial membrane potential (ΔΨM) ( right panel ) measured in crude mitochondria isolated from GSCN of mice fed with ND or HFD. (C) Western blot of CV-ATP5A and CIII-UQRC2 in crude mitochondria isolated from GSCN of mice fed with ND or HFD ( left panel ). Immunoblots reported are representative of three mice per group out of five giving similar results. Densitometric analyses of the immunoreactive bands were performed and normalized to vDAC1 ( right panel ). (D) Western blot analysis of protein carbonyls and Sod2 in crude mitochondria isolated from GSCN of mice fed with ND or HFD. Immunoblots reported are representative of three mice per group out of five giving similar results. Below are reported the densitometric analyses of the immunoreactive bands normalized to TOMM20. (E) mtDNA damage measured by calculating the ratio of long fragment intensity (LFI) and short fragment intensity (SFI) in GSCN of mice fed with ND or HFD. All data are expressed as mean ±S.D. (n = 5 mice/group). Student’s t-test was used to compare ND vs HFD.

    Journal: PLoS ONE

    Article Title: Time-controlled fasting prevents aging-like mitochondrial changes induced by persistent dietary fat overload in skeletal muscle

    doi: 10.1371/journal.pone.0195912

    Figure Lengend Snippet: HFD induces mitochondrial damage and dysfunction in SkM. (A) Basal oxygen consumption measured by a polarographic method in gastrocnemius (GSCN) and tibialis anterior muscle (TA) of mice fed with normal diet (ND) or high fat diet (HFD). (B) Citrate synthase activity ( left panel ) and mitochondrial membrane potential (ΔΨM) ( right panel ) measured in crude mitochondria isolated from GSCN of mice fed with ND or HFD. (C) Western blot of CV-ATP5A and CIII-UQRC2 in crude mitochondria isolated from GSCN of mice fed with ND or HFD ( left panel ). Immunoblots reported are representative of three mice per group out of five giving similar results. Densitometric analyses of the immunoreactive bands were performed and normalized to vDAC1 ( right panel ). (D) Western blot analysis of protein carbonyls and Sod2 in crude mitochondria isolated from GSCN of mice fed with ND or HFD. Immunoblots reported are representative of three mice per group out of five giving similar results. Below are reported the densitometric analyses of the immunoreactive bands normalized to TOMM20. (E) mtDNA damage measured by calculating the ratio of long fragment intensity (LFI) and short fragment intensity (SFI) in GSCN of mice fed with ND or HFD. All data are expressed as mean ±S.D. (n = 5 mice/group). Student’s t-test was used to compare ND vs HFD.

    Article Snippet: Citrate synthase activity was measured by using a Citrate Synthase Activity Assay Kit (Abcam, Cambridge, UK).

    Techniques: Mouse Assay, Activity Assay, Isolation, Western Blot

    SCOT and citrate synthase activities and ATP content. ( a ) SCOT and ( b ) citrate synthase activities as well as ( c ) ATP content of left ventricles of F1B, TO-2, and Bio 14.6 hamsters. Data are mean ± SEM of six animals per group. * P

    Journal: Scientific Reports

    Article Title: Involvement of oxidative modification of proteins related to ATP synthesis in the left ventricles of hamsters with cardiomyopathy

    doi: 10.1038/s41598-017-08546-1

    Figure Lengend Snippet: SCOT and citrate synthase activities and ATP content. ( a ) SCOT and ( b ) citrate synthase activities as well as ( c ) ATP content of left ventricles of F1B, TO-2, and Bio 14.6 hamsters. Data are mean ± SEM of six animals per group. * P

    Article Snippet: Measurement of levels of citrate synthase, SCOT, and ATP Protein samples were prepared according to instructions provided by the manufacturer and the citrate synthase activity in heart tissues was assayed by the Citrate Synthase Activity Assay Kit (Abcam, Cambridge, MA).

    Techniques:

    Correlation of carbonyl contents with the biomolecules and cardiac function. Correlation between LV protein carbonyl contents and ( a ) SCOT activity, ( b ) citrate synthase activity, ( c ) ATP content, ( d ) plasma Troponin T level, ( e ) LV end-diastolic diameter, and ( f ) LV end-systolic diameter. Data are mean ± SEM of six animals per group. Correlation between relative carbonyl levels calculated from the 2D-oxyblot and 2D-DIGE analyses and activities of ( g ) SCOT and ( h ) citrate synthase. Data are mean ± SEM of four animals per group.

    Journal: Scientific Reports

    Article Title: Involvement of oxidative modification of proteins related to ATP synthesis in the left ventricles of hamsters with cardiomyopathy

    doi: 10.1038/s41598-017-08546-1

    Figure Lengend Snippet: Correlation of carbonyl contents with the biomolecules and cardiac function. Correlation between LV protein carbonyl contents and ( a ) SCOT activity, ( b ) citrate synthase activity, ( c ) ATP content, ( d ) plasma Troponin T level, ( e ) LV end-diastolic diameter, and ( f ) LV end-systolic diameter. Data are mean ± SEM of six animals per group. Correlation between relative carbonyl levels calculated from the 2D-oxyblot and 2D-DIGE analyses and activities of ( g ) SCOT and ( h ) citrate synthase. Data are mean ± SEM of four animals per group.

    Article Snippet: Measurement of levels of citrate synthase, SCOT, and ATP Protein samples were prepared according to instructions provided by the manufacturer and the citrate synthase activity in heart tissues was assayed by the Citrate Synthase Activity Assay Kit (Abcam, Cambridge, MA).

    Techniques: Activity Assay

    The effects of exercise on body weight at weeks 1–8 ( n = 20 animals per group) (A), intermittent hypoxia on body weight at weeks 9–10 ( n = 5 animals per group) (B) and exercise on citrate synthase activity ( n = 5 animals per group) (C) . # p

    Journal: Frontiers in Physiology

    Article Title: Exercise Attenuates Intermittent Hypoxia-Induced Cardiac Fibrosis Associated with Sodium-Hydrogen Exchanger-1 in Rats

    doi: 10.3389/fphys.2016.00462

    Figure Lengend Snippet: The effects of exercise on body weight at weeks 1–8 ( n = 20 animals per group) (A), intermittent hypoxia on body weight at weeks 9–10 ( n = 5 animals per group) (B) and exercise on citrate synthase activity ( n = 5 animals per group) (C) . # p

    Article Snippet: The citrate synthase activity of soleus Measurements were obtained using a commercially available citrate synthase activity colorimetric assay kit (BioVision, Milpitas, CA, USA), according to the manufacturer's instructions.

    Techniques: Activity Assay

    CSL-111 treatment normalizes hypothalamic inflammation and mitochondrial function markers in chow- fed apoA-I −/− mice. Plasma total cholesterol levels (A) and lipoprotein fractions (B) of apoA-I +/+ and apoA-I −/− mice after daily treatment with CSL-111 at 150 mg/kg for 7 days (n = 6 mice per group, age 7 months). CSL-111 normalizes hypothalamic inflammation (C) and reverses mitochondrial changes by enhancing citrate synthase activity (D) and normalizing mitochondrial function markers (E) in apoA-I −/− mice. Body composition analysis revealed reduced fat mass in apoA-I +/+ (white hatched bars) and apoA-I −/− mice (red hatched bars) treated with CSL-111 compared with vehicle-treated groups (F). The changes in body composition were due to reduced visceral fat mass in apoA-I +/+ mice (white hatched bars) (G) and due to reduced hepatic fat content in apoA-I −/− mice (red hatched bars) (H). Representative pictures of H E-stained hepatic sections from WT and apoA-I −/− mice revealed accumulation of hepatic lipids in the form of lipid droplets within the hepatocytes in vehicle-treated apoA-I −/− mice (40× magnification), which normalized in the CSL-111 treatment group. CSL-111 treatment normalized cholesterol (J) and triglyceride (K) levels in the CNS of apoA-I −/− mice. Data are expressed as mean ± SEM. * P

    Journal: Journal of Lipid Research

    Article Title: Circulating HDL levels control hypothalamic astrogliosis via apoA-I

    doi: 10.1194/jlr.M085456

    Figure Lengend Snippet: CSL-111 treatment normalizes hypothalamic inflammation and mitochondrial function markers in chow- fed apoA-I −/− mice. Plasma total cholesterol levels (A) and lipoprotein fractions (B) of apoA-I +/+ and apoA-I −/− mice after daily treatment with CSL-111 at 150 mg/kg for 7 days (n = 6 mice per group, age 7 months). CSL-111 normalizes hypothalamic inflammation (C) and reverses mitochondrial changes by enhancing citrate synthase activity (D) and normalizing mitochondrial function markers (E) in apoA-I −/− mice. Body composition analysis revealed reduced fat mass in apoA-I +/+ (white hatched bars) and apoA-I −/− mice (red hatched bars) treated with CSL-111 compared with vehicle-treated groups (F). The changes in body composition were due to reduced visceral fat mass in apoA-I +/+ mice (white hatched bars) (G) and due to reduced hepatic fat content in apoA-I −/− mice (red hatched bars) (H). Representative pictures of H E-stained hepatic sections from WT and apoA-I −/− mice revealed accumulation of hepatic lipids in the form of lipid droplets within the hepatocytes in vehicle-treated apoA-I −/− mice (40× magnification), which normalized in the CSL-111 treatment group. CSL-111 treatment normalized cholesterol (J) and triglyceride (K) levels in the CNS of apoA-I −/− mice. Data are expressed as mean ± SEM. * P

    Article Snippet: Brain cortex tissue was snap-frozen for determination of citrate synthase activity by a colorimetric assay kit (K318-100; Biovision, Germany).

    Techniques: Mouse Assay, Activity Assay, Staining

    apoA-I directly enhances cellular respiration by increasing mitochondrial activity in primary astrocytes. Cellular respiration and respiration after interference of mitochondrial pathways with specific inhibitors was analyzed in astrocytes incubated with BSA or apoA-I (20 μg/ml overnight) using an XF24 extracellular flux analyzer as described in the Materials and Methods. A: OCR over time in primary astrocytes treated with apoA-I (filled circles) and BSA (open circles). B: Scheme defining the oxygen-consuming processes of a cell. C: Cellular respiration was dissected into mitochondrial (green) and nonmitochondrial (gray) respiration using the electron transfer chain inhibitors, rotenone (R; 25 μM) and antimycin A (AA; 25 μM). D: Mitochondrial respiration was further dissected into proton leak (dark gray) and ATP-linked respiration (red) using the ATP-synthase inhibitor, oligomycin (oligo; 20 μg/ml). E: Maximal respiration (dark blue) of astrocytes after addition of the chemical uncoupler, FCCP (10 μM), and after subtracting nonmitochondrial respiration rates. The portion of spare respiratory capacity (light blue) was determined by subtracting basal respiration from maximal respiration rates. F: Mitochondrial coupling efficiencies were calculated as the fraction of mitochondrial oxygen consumption that is sensitive to oligomycin, reflecting the fraction used to drive ATP synthesis. AU, arbitrary units. All data are expressed as mean ± SEM of five replicates run in parallel. * P

    Journal: Journal of Lipid Research

    Article Title: Circulating HDL levels control hypothalamic astrogliosis via apoA-I

    doi: 10.1194/jlr.M085456

    Figure Lengend Snippet: apoA-I directly enhances cellular respiration by increasing mitochondrial activity in primary astrocytes. Cellular respiration and respiration after interference of mitochondrial pathways with specific inhibitors was analyzed in astrocytes incubated with BSA or apoA-I (20 μg/ml overnight) using an XF24 extracellular flux analyzer as described in the Materials and Methods. A: OCR over time in primary astrocytes treated with apoA-I (filled circles) and BSA (open circles). B: Scheme defining the oxygen-consuming processes of a cell. C: Cellular respiration was dissected into mitochondrial (green) and nonmitochondrial (gray) respiration using the electron transfer chain inhibitors, rotenone (R; 25 μM) and antimycin A (AA; 25 μM). D: Mitochondrial respiration was further dissected into proton leak (dark gray) and ATP-linked respiration (red) using the ATP-synthase inhibitor, oligomycin (oligo; 20 μg/ml). E: Maximal respiration (dark blue) of astrocytes after addition of the chemical uncoupler, FCCP (10 μM), and after subtracting nonmitochondrial respiration rates. The portion of spare respiratory capacity (light blue) was determined by subtracting basal respiration from maximal respiration rates. F: Mitochondrial coupling efficiencies were calculated as the fraction of mitochondrial oxygen consumption that is sensitive to oligomycin, reflecting the fraction used to drive ATP synthesis. AU, arbitrary units. All data are expressed as mean ± SEM of five replicates run in parallel. * P

    Article Snippet: Brain cortex tissue was snap-frozen for determination of citrate synthase activity by a colorimetric assay kit (K318-100; Biovision, Germany).

    Techniques: Activity Assay, Incubation

    mtCaMKII activates TCA cycle dehydrogenases and augments resting NADH. a Pyruvate dehydrogenase (PDH) activity in isolated mitochondria from WT ( n = 7) and mtCaMKII ( n = 9) hearts. b TCA cycle enzyme activities in permeabilized mitochondria from WT ( n = 7) and mtCaMKII ( n = 9) hearts, CS: citrate synthase, IDH: isocitrate dehydrogenase, α-KGDH: α-ketoglutarate dehydrogenase, SL: succinate-CoA ligase, SDH: succinate dehydrogenase, MDH: malate dehydrogenase. c Representative NADH imaging of isolated ventricular myocytes before pacing (calibration bar = 50 μm). d Quantification of NADH autofluorescence in isolated ventricular myocytes before and after pacing from WT ( n = 29 myocytes from two hearts) and mtCaMKII ( n = 52 myocytes from three hearts) hearts. Maximum (100%) NADH was measured in the presence of NaCN, and minimum (0%) in the presence of FCCP. Cells from the same genotype were paired in comparisons of before and after pacing. e Quantification of NAD + and NADH in hearts from WT ( n = 4) and mtCaMKII ( n = 4) mice. f Western blot and g summary data for acetylated proteins normalized to VDAC1 in isolated mitochondria from WT ( n = 4) and mtCaMKII ( n = 4) hearts. Data are represented as mean ± SEM, significance was determined using a two-tailed Student’s t test. **** P

    Journal: Nature Communications

    Article Title: Mitochondrial CaMKII causes adverse metabolic reprogramming and dilated cardiomyopathy

    doi: 10.1038/s41467-020-18165-6

    Figure Lengend Snippet: mtCaMKII activates TCA cycle dehydrogenases and augments resting NADH. a Pyruvate dehydrogenase (PDH) activity in isolated mitochondria from WT ( n = 7) and mtCaMKII ( n = 9) hearts. b TCA cycle enzyme activities in permeabilized mitochondria from WT ( n = 7) and mtCaMKII ( n = 9) hearts, CS: citrate synthase, IDH: isocitrate dehydrogenase, α-KGDH: α-ketoglutarate dehydrogenase, SL: succinate-CoA ligase, SDH: succinate dehydrogenase, MDH: malate dehydrogenase. c Representative NADH imaging of isolated ventricular myocytes before pacing (calibration bar = 50 μm). d Quantification of NADH autofluorescence in isolated ventricular myocytes before and after pacing from WT ( n = 29 myocytes from two hearts) and mtCaMKII ( n = 52 myocytes from three hearts) hearts. Maximum (100%) NADH was measured in the presence of NaCN, and minimum (0%) in the presence of FCCP. Cells from the same genotype were paired in comparisons of before and after pacing. e Quantification of NAD + and NADH in hearts from WT ( n = 4) and mtCaMKII ( n = 4) mice. f Western blot and g summary data for acetylated proteins normalized to VDAC1 in isolated mitochondria from WT ( n = 4) and mtCaMKII ( n = 4) hearts. Data are represented as mean ± SEM, significance was determined using a two-tailed Student’s t test. **** P

    Article Snippet: Citrate synthase (CS) activity was measured using a MitoCheck Citrate Synthase Activity Assay Kit (Cayman Chemicals).

    Techniques: Activity Assay, Isolation, Imaging, Mouse Assay, Western Blot, Two Tailed Test

    Influence of SIRT3 depletion on mitochondrial activity. A) Respiration rates in C2C12- LucshRNA (shCTL) and C2C12-SIRT3shRNA (shSIRT3) at cell proliferation (P), cell confluence (C) and 3 days of differentiation (D3). Values are normalized relatively to total protein levels. B) Citrate synthase, complex II and cytochrome c oxidase maximal activities in shCTL and shSIRT3 cells at cell proliferation (P), cell confluence (C) and 3 days of differentiation (D3). Results are expressed as mean ± SD from five independent experiments. ANOVA main effect: #P

    Journal: PLoS ONE

    Article Title: SIRT3, a Mitochondrial NAD+-Dependent Deacetylase, Is Involved in the Regulation of Myoblast Differentiation

    doi: 10.1371/journal.pone.0114388

    Figure Lengend Snippet: Influence of SIRT3 depletion on mitochondrial activity. A) Respiration rates in C2C12- LucshRNA (shCTL) and C2C12-SIRT3shRNA (shSIRT3) at cell proliferation (P), cell confluence (C) and 3 days of differentiation (D3). Values are normalized relatively to total protein levels. B) Citrate synthase, complex II and cytochrome c oxidase maximal activities in shCTL and shSIRT3 cells at cell proliferation (P), cell confluence (C) and 3 days of differentiation (D3). Results are expressed as mean ± SD from five independent experiments. ANOVA main effect: #P

    Article Snippet: In order to test the influence of SIRT3 on mitochondrial biogenesis, we measured the expression of markers of the mitochondrial mass: PGC-1α, a major regulator of mitochondrial biogenesis and citrate synthase ( ).

    Techniques: Activity Assay