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  • 99
    Millipore nad nadh assay nad
    Analysis of respiratory activity and ATP synthesis with pyruvate in HT29, HT29-dx and HMM cells. ( A ) The cells were incubated in fresh medium in the absence or in the presence of pyruvate (pyr) 25 mM for 24 h. The cells were lysed, the mitochondrial fractions were isolated and the rate of reduction of cytochrome c was measured. ( B ) ATP was measured in triplicate in the mitochondrial extracts by a chemiluminescence-based assay. ( C ) intracellular <t>NAD</t> + <t>/NADH</t> ratio was measured in triplicate in the whole cells by a chemiluminescence-based assay. Measurements were performed in triplicate and data are presented as means ± SEM ( n = 3). ( A , B ) HT29-dx and HMM cells in the absence of pyr versus HT29 and HMM cells in the absence of pyr (ctrl) *** p
    Nad Nadh Assay Nad, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam nad nadh nad nadh
    Systemic exposure to 18% oxygen regulates levels of <t>NAD</t> +, <t>NADH,</t> lactate, LDH and SUR2A in the heart. Bar graphs depicting PO 2 (O 2 ), PCO 2 (CO 2 ) and haematocrit (HCT) in the blood as well as ATP (ATP), lactate (Lactate), LDH, total NAD (NADt), NADH (NADH), NAD + (NAD +) and SUR2A (SUR2A) in the myocardial tissue; inset in SUR2A bar graph represents original Western blots for SUR2A under labelled conditions. Each bar is a mean ± SEM (n = 3–5). *P
    Nad Nadh Nad Nadh, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam nicotinamide adenine dinucleotide nad levels
    Intracellular total <t>NAD</t> levels in platelet lysates. Total NAD was measured in platelet lysates during storage of PC. Dotted lines indicate when addition of NR or vehicle solution was performed. Samples were collected on Days 1, 5, 7, 12, 19, and 23 from PC supplemented with NR ( ○ ) or control (●). Data are shown as mean with SD (n = 6), ****p
    Nicotinamide Adenine Dinucleotide Nad Levels, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Millipore nadh nad
    Effect of MeHg on the production of <t>NAD(H).</t> Undifferentiated PC12 cells were treated with 0μM (white) or 2μM (black) MeHg for 60 min, and then the amounts of NAD + (A) and <t>NADH</t> (B) were measured by spectrophotometry using an NAD
    Nadh Nad, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore β nicotinamide adenine dinucleotide nadh
    Effect of MeHg on the production of <t>NAD(H).</t> Undifferentiated PC12 cells were treated with 0μM (white) or 2μM (black) MeHg for 60 min, and then the amounts of NAD + (A) and <t>NADH</t> (B) were measured by spectrophotometry using an NAD
    β Nicotinamide Adenine Dinucleotide Nadh, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 60 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Millipore nad nadh kit
    Effect of MeHg on the production of <t>NAD(H).</t> Undifferentiated PC12 cells were treated with 0μM (white) or 2μM (black) MeHg for 60 min, and then the amounts of NAD + (A) and <t>NADH</t> (B) were measured by spectrophotometry using an NAD
    Nad Nadh Kit, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Analysis of respiratory activity and ATP synthesis with pyruvate in HT29, HT29-dx and HMM cells. ( A ) The cells were incubated in fresh medium in the absence or in the presence of pyruvate (pyr) 25 mM for 24 h. The cells were lysed, the mitochondrial fractions were isolated and the rate of reduction of cytochrome c was measured. ( B ) ATP was measured in triplicate in the mitochondrial extracts by a chemiluminescence-based assay. ( C ) intracellular NAD + /NADH ratio was measured in triplicate in the whole cells by a chemiluminescence-based assay. Measurements were performed in triplicate and data are presented as means ± SEM ( n = 3). ( A , B ) HT29-dx and HMM cells in the absence of pyr versus HT29 and HMM cells in the absence of pyr (ctrl) *** p

    Journal: International Journal of Molecular Sciences

    Article Title: Pyruvate Treatment Restores the Effectiveness of Chemotherapeutic Agents in Human Colon Adenocarcinoma and Pleural Mesothelioma Cells

    doi: 10.3390/ijms19113550

    Figure Lengend Snippet: Analysis of respiratory activity and ATP synthesis with pyruvate in HT29, HT29-dx and HMM cells. ( A ) The cells were incubated in fresh medium in the absence or in the presence of pyruvate (pyr) 25 mM for 24 h. The cells were lysed, the mitochondrial fractions were isolated and the rate of reduction of cytochrome c was measured. ( B ) ATP was measured in triplicate in the mitochondrial extracts by a chemiluminescence-based assay. ( C ) intracellular NAD + /NADH ratio was measured in triplicate in the whole cells by a chemiluminescence-based assay. Measurements were performed in triplicate and data are presented as means ± SEM ( n = 3). ( A , B ) HT29-dx and HMM cells in the absence of pyr versus HT29 and HMM cells in the absence of pyr (ctrl) *** p

    Article Snippet: NAD+ /NADH Assay NAD+ and NADH were measured using the NAD+ /NADH Quantification Colorimetric Kit from Sigma-Aldrich (Milan, Italy), which allows determination of NAD+ , NADH and their ratio, without the requirement to purification steps.

    Techniques: Activity Assay, Incubation, Isolation, Chemiluminescence Immunoassay

    ATGL is required for the β-adrenergic induction of SIRT1 and PGC1-α activity. A and B : Inhibition of ATGL blocks cAMP induction of SIRT1 activity. Primary hepatocytes were treated with or without 1 mmol/L cAMP analog (8-bromoadenosine 3′,5′-cyclic monophosphate) for 10 min after a 1-h pretreatment with 2 μmol/L ATGL inhibitor BEL ( A ) or 30 μmol/L ATGL inhibitor Astat ( B ). SIRT1 activity was determined by fluorometric kinetic assay from hepatocyte nuclear lysates ( n = 3). C : SIRT1 activity is inhibited by a PKA inhibitor. Primary hepatocytes were treated with cAMP analog following a 1-h preincubation with 30 μmol/L H89 ( n = 3). D : Inhibition of ATGL blocks cAMP induction of PGC-1α reporter expression. PGC-1α reporter was transfected into primary hepatocytes, and reporter activity was determined by dual-luciferase reporter assay ( n = 3). E : NAD + levels in mouse liver tissue treated with indicated adenoviruses for 7 days as determined by enzyme-based NAD/NADH quantification kit ( n = 4). F : p38-MAPK phosphorylation in livers of mice with overexpressed or knocked down ATGL. Graphs reflect fold change over control in densitometry of acetylated proteins, normalized to total protein ( n = 3). * P

    Journal: Diabetes

    Article Title: ATGL-Catalyzed Lipolysis Regulates SIRT1 to Control PGC-1α/PPAR-α Signaling

    doi: 10.2337/db14-0325

    Figure Lengend Snippet: ATGL is required for the β-adrenergic induction of SIRT1 and PGC1-α activity. A and B : Inhibition of ATGL blocks cAMP induction of SIRT1 activity. Primary hepatocytes were treated with or without 1 mmol/L cAMP analog (8-bromoadenosine 3′,5′-cyclic monophosphate) for 10 min after a 1-h pretreatment with 2 μmol/L ATGL inhibitor BEL ( A ) or 30 μmol/L ATGL inhibitor Astat ( B ). SIRT1 activity was determined by fluorometric kinetic assay from hepatocyte nuclear lysates ( n = 3). C : SIRT1 activity is inhibited by a PKA inhibitor. Primary hepatocytes were treated with cAMP analog following a 1-h preincubation with 30 μmol/L H89 ( n = 3). D : Inhibition of ATGL blocks cAMP induction of PGC-1α reporter expression. PGC-1α reporter was transfected into primary hepatocytes, and reporter activity was determined by dual-luciferase reporter assay ( n = 3). E : NAD + levels in mouse liver tissue treated with indicated adenoviruses for 7 days as determined by enzyme-based NAD/NADH quantification kit ( n = 4). F : p38-MAPK phosphorylation in livers of mice with overexpressed or knocked down ATGL. Graphs reflect fold change over control in densitometry of acetylated proteins, normalized to total protein ( n = 3). * P

    Article Snippet: NAD+ and NADH Measurements NAD+ and NADH concentrations were determined using a colorimetric, enzyme-based NAD+ /NADH quantification kit (Sigma-Aldrich) according to the manufacturer’s instructions.

    Techniques: Activity Assay, Inhibition, Kinetic Assay, Pyrolysis Gas Chromatography, Expressing, Transfection, Luciferase, Reporter Assay, Mouse Assay

    Niacin prevents m-HCD driving senescent phenotype in sAT of newborn mice A. , B. Analyses of data from the publicly available GEO data set revealed a detectable expression of genes related to NAD + synthesis from niacin (Preiss-Handler pathway) in adult white adipose tissue (ID: GDS6247) A. and proliferating adipocytes precursors (ID: GDS5824) B. of mice fed with NCD or HCD. C. NAD + /NADH ratio in sAT of newborn mice supplemented or not with niacin. D. Measurement of the mRNA expression of Preiss-Handler pathway and HCAR2 genes in sAT of newborn mice supplemented or not with niacin. E. Fold change in mtDNA/nDNA ratio ( left panel ) and citrate synthase (CS) activity ( right panel ) detected in sAT of newborn mice supplemented or not with niacin. F. Protein levels of PARP1, SIRT1 and acetylated FoxO1 form in sAT of newborn mice supplemented or not with niacin ( left panel ), densitometric analysis of PARP1/SIRT1 ( central panel ) and acetyl FoxO1 protein ratio ( right panel ). G. , H. Protein levels of mitochondrial OxPHOS subunits detected on crude mitochondria G. , mtDNA-encoded and nDNA-encoded genes expression H. in sAT of newborn mice born from mothers supplemented or not with niacin. I. , J. Analysis of senescence I. and inflammatory J. markers in sAT of newborn mice supplemented or not with niacin. K. Glucose, cholesterol and triglyceride levels detected in blood of mothers 12 weeks after NCD and HCD. β-Actin served as loading control. All data are expressed as mean ±S.D. ( n = 6 female newborn mice/group; * p

    Journal: Oncotarget

    Article Title: Maternal high calorie diet induces mitochondrial dysfunction and senescence phenotype in subcutaneous fat of newborn mice

    doi: 10.18632/oncotarget.19948

    Figure Lengend Snippet: Niacin prevents m-HCD driving senescent phenotype in sAT of newborn mice A. , B. Analyses of data from the publicly available GEO data set revealed a detectable expression of genes related to NAD + synthesis from niacin (Preiss-Handler pathway) in adult white adipose tissue (ID: GDS6247) A. and proliferating adipocytes precursors (ID: GDS5824) B. of mice fed with NCD or HCD. C. NAD + /NADH ratio in sAT of newborn mice supplemented or not with niacin. D. Measurement of the mRNA expression of Preiss-Handler pathway and HCAR2 genes in sAT of newborn mice supplemented or not with niacin. E. Fold change in mtDNA/nDNA ratio ( left panel ) and citrate synthase (CS) activity ( right panel ) detected in sAT of newborn mice supplemented or not with niacin. F. Protein levels of PARP1, SIRT1 and acetylated FoxO1 form in sAT of newborn mice supplemented or not with niacin ( left panel ), densitometric analysis of PARP1/SIRT1 ( central panel ) and acetyl FoxO1 protein ratio ( right panel ). G. , H. Protein levels of mitochondrial OxPHOS subunits detected on crude mitochondria G. , mtDNA-encoded and nDNA-encoded genes expression H. in sAT of newborn mice born from mothers supplemented or not with niacin. I. , J. Analysis of senescence I. and inflammatory J. markers in sAT of newborn mice supplemented or not with niacin. K. Glucose, cholesterol and triglyceride levels detected in blood of mothers 12 weeks after NCD and HCD. β-Actin served as loading control. All data are expressed as mean ±S.D. ( n = 6 female newborn mice/group; * p

    Article Snippet: NAD+ /NADH ratio NAD+ /NADH ratio was detected in total homogenates by using NAD+ /NADH Quantitation Kit according to manufacturer's instruction (Sigma-Aldrich).

    Techniques: Mouse Assay, Expressing, Activity Assay

    Maternal-HCD induces NAD + decrease, mitochondrial dysfunction and SASP in sAT of newborn mice A. Cytofluorimetric analyses of mitochondrial membrane potential ( left panel ) and enzymatic determination of citrate synthase activity (CS) ( right panel ) in sAT of newborn mice. B. , C. mRNA expression of mtDNA-encoded and nDNA-encoded genes B. and protein levels C. of mitochondrial OxPHOS subunits in sAT of newborn mice. D. SOD2 and UCP1 protein levels ( left panel ) and their densitometric analyses ( right panel ) in sAT crude mitochondria of newborn mice. E. Protein carbonylation in sAT crude mitochondria of newborn mice. F. Oxygen consumption and respiratory control index in sAT crude mitochondria of newborn mice. G. NAD + /NADH ratio measured in total homogenates from sAT of newborn mice. H. Protein levels of PARP1, SIRT1, FoxO1 and its acetylated form ( left panel ), and densitometric analysis of PARP1/SIRT1 protein ratio ( right panel ) detected in sAT of newborn mice. I. Cytoplasmic levels of FoxO1 in sAT of newborn mice. 3T3-L1 lysates served as control of FoxO1 antibody specificity. J. Schematic representation of the m-HCD impact on mitochondrial adaptations and molecular responses in newborn mice. β-Actin and vDAC1 served as loading control. For figures D and E the same loading control was reported. All data are expressed as mean ±S.D. ( n = 6 female newborn mice/group; * p

    Journal: Oncotarget

    Article Title: Maternal high calorie diet induces mitochondrial dysfunction and senescence phenotype in subcutaneous fat of newborn mice

    doi: 10.18632/oncotarget.19948

    Figure Lengend Snippet: Maternal-HCD induces NAD + decrease, mitochondrial dysfunction and SASP in sAT of newborn mice A. Cytofluorimetric analyses of mitochondrial membrane potential ( left panel ) and enzymatic determination of citrate synthase activity (CS) ( right panel ) in sAT of newborn mice. B. , C. mRNA expression of mtDNA-encoded and nDNA-encoded genes B. and protein levels C. of mitochondrial OxPHOS subunits in sAT of newborn mice. D. SOD2 and UCP1 protein levels ( left panel ) and their densitometric analyses ( right panel ) in sAT crude mitochondria of newborn mice. E. Protein carbonylation in sAT crude mitochondria of newborn mice. F. Oxygen consumption and respiratory control index in sAT crude mitochondria of newborn mice. G. NAD + /NADH ratio measured in total homogenates from sAT of newborn mice. H. Protein levels of PARP1, SIRT1, FoxO1 and its acetylated form ( left panel ), and densitometric analysis of PARP1/SIRT1 protein ratio ( right panel ) detected in sAT of newborn mice. I. Cytoplasmic levels of FoxO1 in sAT of newborn mice. 3T3-L1 lysates served as control of FoxO1 antibody specificity. J. Schematic representation of the m-HCD impact on mitochondrial adaptations and molecular responses in newborn mice. β-Actin and vDAC1 served as loading control. For figures D and E the same loading control was reported. All data are expressed as mean ±S.D. ( n = 6 female newborn mice/group; * p

    Article Snippet: NAD+ /NADH ratio NAD+ /NADH ratio was detected in total homogenates by using NAD+ /NADH Quantitation Kit according to manufacturer's instruction (Sigma-Aldrich).

    Techniques: Mouse Assay, Activity Assay, Expressing

    HCD induced mitochondrial dysfunction in on sAT of adult C57BL/6J female mice A. Analyses of data from the publicly available GEO data set (ID: GDS3102) revealed a down-regulation of mitochondrial OxPHOS subunits and TCA enzymes in the sAT of old (28 mo.) versus young (4 mo.) rats. Oppositely, inflammatory cytokines levels were higher in old than young rats. Calorie restriction (CR) significantly improved OxPHOS subunits and inflammatory cytokines gene transcription. Gene expression profile was obtained calculating the fold change of gene expression profile provided by gene expression omnibus (ID: GDS3102). For details related to gene included in our analyses, see Experimental Procedures section. B. Oxygen consumption and mitochondrial membrane potential measured in sAT-derived crude mitochondria of adult female mice 6 and 12 weeks after normal calorie diet (NCD) and high calorie diet (HCD). C. Mitochondrial mass evaluated by analysing the protein levels of mitochondrial TOMM20 in total sAT homogenates 12 weeks after NCD and HCD. Boxes reported below show the percentage of TOMM20 protein in total fraction ( left panel ). Fold change in citrate synthase (CS) activity and mtDNA/nDNA ratio detected in sAT 12 weeks after NCD and HCD ( right panel ). D. Measurement of the mRNA expression of mtDNA-encoded genes in sAT 12 weeks after NCD and HCD. E. NAD + /NADH ratio measured in total homogenates of sAT of mothers 12 weeks after NCD and HCD. F. PARP1 and SIRT1 protein levels and their densitometric analyses ( right panel ) in total homogenates of sAT 12 weeks after NCD and HCD. G. Senescence-associated β-galactosidase activity ( upper panel ) and mRNA expression of inflammatory cytokines ( lower panel ) in sAT 12 weeks after NCD and HCD. Tubulin served as loading control. All data are expressed as mean ±S.D. ( n = 4 female adult mice/group; * p

    Journal: Oncotarget

    Article Title: Maternal high calorie diet induces mitochondrial dysfunction and senescence phenotype in subcutaneous fat of newborn mice

    doi: 10.18632/oncotarget.19948

    Figure Lengend Snippet: HCD induced mitochondrial dysfunction in on sAT of adult C57BL/6J female mice A. Analyses of data from the publicly available GEO data set (ID: GDS3102) revealed a down-regulation of mitochondrial OxPHOS subunits and TCA enzymes in the sAT of old (28 mo.) versus young (4 mo.) rats. Oppositely, inflammatory cytokines levels were higher in old than young rats. Calorie restriction (CR) significantly improved OxPHOS subunits and inflammatory cytokines gene transcription. Gene expression profile was obtained calculating the fold change of gene expression profile provided by gene expression omnibus (ID: GDS3102). For details related to gene included in our analyses, see Experimental Procedures section. B. Oxygen consumption and mitochondrial membrane potential measured in sAT-derived crude mitochondria of adult female mice 6 and 12 weeks after normal calorie diet (NCD) and high calorie diet (HCD). C. Mitochondrial mass evaluated by analysing the protein levels of mitochondrial TOMM20 in total sAT homogenates 12 weeks after NCD and HCD. Boxes reported below show the percentage of TOMM20 protein in total fraction ( left panel ). Fold change in citrate synthase (CS) activity and mtDNA/nDNA ratio detected in sAT 12 weeks after NCD and HCD ( right panel ). D. Measurement of the mRNA expression of mtDNA-encoded genes in sAT 12 weeks after NCD and HCD. E. NAD + /NADH ratio measured in total homogenates of sAT of mothers 12 weeks after NCD and HCD. F. PARP1 and SIRT1 protein levels and their densitometric analyses ( right panel ) in total homogenates of sAT 12 weeks after NCD and HCD. G. Senescence-associated β-galactosidase activity ( upper panel ) and mRNA expression of inflammatory cytokines ( lower panel ) in sAT 12 weeks after NCD and HCD. Tubulin served as loading control. All data are expressed as mean ±S.D. ( n = 4 female adult mice/group; * p

    Article Snippet: NAD+ /NADH ratio NAD+ /NADH ratio was detected in total homogenates by using NAD+ /NADH Quantitation Kit according to manufacturer's instruction (Sigma-Aldrich).

    Techniques: Mouse Assay, Expressing, Derivative Assay, Activity Assay

    Effect of WY-14643 administration in NAMPT protein expression and NAD + /NADH levels. (a) Western blot and densitometric analysis of NAMPT. (b) Photometric analysis of NAD + /NADH levels in steatotic livers after 24 hours of reperfusion. Sham: anesthesia and laparotomy, IR: 60 min partial ischemia and 24 h of reperfusion, and WY-14643 + IR: iv administration of WY-14643 (10 mg/kg) 1 hour before IR. ∗ P

    Journal: BioMed Research International

    Article Title: PPARα Agonist WY-14643 Induces SIRT1 Activity in Rat Fatty Liver Ischemia-Reperfusion Injury

    doi: 10.1155/2015/894679

    Figure Lengend Snippet: Effect of WY-14643 administration in NAMPT protein expression and NAD + /NADH levels. (a) Western blot and densitometric analysis of NAMPT. (b) Photometric analysis of NAD + /NADH levels in steatotic livers after 24 hours of reperfusion. Sham: anesthesia and laparotomy, IR: 60 min partial ischemia and 24 h of reperfusion, and WY-14643 + IR: iv administration of WY-14643 (10 mg/kg) 1 hour before IR. ∗ P

    Article Snippet: NAD+ /NADH Determination Hepatic NAD+ /NADH levels were quantified with a commercially available kit (MAK037, Sigma Chemical, St. Louis, MO, United States) according to the manufacturer's instructions.

    Techniques: Expressing, Western Blot

    Constraints-based Modeling predicts disruption of redox homeostasis and rewiring of metabolic network for compensation. a Core network representation of C. violaceum metabolism (iDB149) with tryptophan, violacein pathway (using Escher; https://escher.github.io/ ) and tailored biomass composition. b Reconstruction statistics and subsystem classification. c - e NADH and NAD experimental values attained for the three different strains using three different substrates – Glucose, Pyruvate and Succinate. Mean ± S.D. for triplicate samples represented

    Journal: BMC Systems Biology

    Article Title: A scalable metabolite supplementation strategy against antibiotic resistant pathogen Chromobacterium violaceum induced by NAD+/NADH+ imbalance

    doi: 10.1186/s12918-017-0427-z

    Figure Lengend Snippet: Constraints-based Modeling predicts disruption of redox homeostasis and rewiring of metabolic network for compensation. a Core network representation of C. violaceum metabolism (iDB149) with tryptophan, violacein pathway (using Escher; https://escher.github.io/ ) and tailored biomass composition. b Reconstruction statistics and subsystem classification. c - e NADH and NAD experimental values attained for the three different strains using three different substrates – Glucose, Pyruvate and Succinate. Mean ± S.D. for triplicate samples represented

    Article Snippet: NADH and NAD measurements NAD/NADH levels of C. violaceum cultures (WT, ChlR and StrpR) were measured using a commercially available kit (MAK037, Sigma Chemical, St. Louis, MO, United States) according to the manufacturer’s instructions (Fig. ).

    Techniques:

    Alteration in carbon metabolism upon polyQ intoxication. a Growth of transformed yeasts was monitored after 8 days of incubation. The pica phenotype of Q 56 -YFP is enhanced when BY4741 cells are grown on SD media containing 2 % sorbitol or 2 % glycerol. The pica phenotype is unchanged if cells are grown on 2 % galactose as a carbon source compared to glucose. Upon respiratory chain inhibition by adding oligomycin no growth is observable after pQ56 transformation. The scale bar represents 10 mm. b Yeast were resuspended to an OD 595 = 150 and [U- 13 C 6 ]-glucose was added at time point 0. Spectra were recorded for 3 h by NMR. The kinetics of glucose consumption are based on the peak at 75.8 ppm and ethanol production based on the peak at 16.7 ppm in pQ0- (black square) and pQ56- (red circle) transformed yeasts. The kinetics of pyruvate accumulation are based on the peaks at 169.9 ppm and carbonate production quantified based on the peak at 160.3 ppm in pQ0- (black square) and pQ56- (red circle) transformed yeasts. The chemical shift positions are assigned in Additional file 13 and Additional file 14 F and G. c ATP-level of Q 0 -YFP, Q 30 -YFP and Q 56 -YFP yeasts was determined using a luciferase coupled assay. Means and SEM are depicted. Six biological replicates were analyzed. The detected differences are not significant. d NADH-level of Q 0 -YFP, Q 30 -YFP and Q 56 -YFP yeasts. The detected differences are not significant. e ) MitoTracker staining in Q 56 -YFP and Q 0 -YFP expressing cells. Expression time is not changed between samples. Scale bar represents 5 μm for MitoTracker. Expression pattern of Cox4p and Om45p fused to GFP. Exposure time is not changed between Q 56 -mCherry and Q 0 -mCherry producing cells. Scale bar represents 10 μm for Cox4-GFP and Om45-GFP. Quantification was carried out as described in the methods section. Mean fluorescence per pixel is plotted with SD

    Journal: BMC Genomics

    Article Title: Polyglutamine toxicity in yeast induces metabolic alterations and mitochondrial defects

    doi: 10.1186/s12864-015-1831-7

    Figure Lengend Snippet: Alteration in carbon metabolism upon polyQ intoxication. a Growth of transformed yeasts was monitored after 8 days of incubation. The pica phenotype of Q 56 -YFP is enhanced when BY4741 cells are grown on SD media containing 2 % sorbitol or 2 % glycerol. The pica phenotype is unchanged if cells are grown on 2 % galactose as a carbon source compared to glucose. Upon respiratory chain inhibition by adding oligomycin no growth is observable after pQ56 transformation. The scale bar represents 10 mm. b Yeast were resuspended to an OD 595 = 150 and [U- 13 C 6 ]-glucose was added at time point 0. Spectra were recorded for 3 h by NMR. The kinetics of glucose consumption are based on the peak at 75.8 ppm and ethanol production based on the peak at 16.7 ppm in pQ0- (black square) and pQ56- (red circle) transformed yeasts. The kinetics of pyruvate accumulation are based on the peaks at 169.9 ppm and carbonate production quantified based on the peak at 160.3 ppm in pQ0- (black square) and pQ56- (red circle) transformed yeasts. The chemical shift positions are assigned in Additional file 13 and Additional file 14 F and G. c ATP-level of Q 0 -YFP, Q 30 -YFP and Q 56 -YFP yeasts was determined using a luciferase coupled assay. Means and SEM are depicted. Six biological replicates were analyzed. The detected differences are not significant. d NADH-level of Q 0 -YFP, Q 30 -YFP and Q 56 -YFP yeasts. The detected differences are not significant. e ) MitoTracker staining in Q 56 -YFP and Q 0 -YFP expressing cells. Expression time is not changed between samples. Scale bar represents 5 μm for MitoTracker. Expression pattern of Cox4p and Om45p fused to GFP. Exposure time is not changed between Q 56 -mCherry and Q 0 -mCherry producing cells. Scale bar represents 10 μm for Cox4-GFP and Om45-GFP. Quantification was carried out as described in the methods section. Mean fluorescence per pixel is plotted with SD

    Article Snippet: NAD + /NADH levels were determined using a commercial available NAD/NADH Quantification Kit (Sigma-Aldrich, St. Louis, MO, USA).

    Techniques: Transformation Assay, Incubation, IF-cells, Inhibition, Nuclear Magnetic Resonance, Luciferase, Staining, Expressing, Fluorescence

    Influence of maternal fat supplementation on skeletal muscle enzymatic activity. (A) Enzymatic activities of total lactate dehydrogenase (LDH) and isocitrate dehydrogenase (ICDH) and their ratios in the bicep femoris . (B) Muscular concentrations and ratio of nicotinamide adenine dinucleotide (NADH) as well as its reduced form NAD + on 7-day old offspring exposed to a control (white; n = 7) or high fat (black; n = 7) maternal diet throughout gestation. Bar graphs illustrate means ± SEM (#p

    Journal: BMC Physiology

    Article Title: Impact of maternal dietary fat supplementation during gestation upon skeletal muscle in neonatal pigs

    doi: 10.1186/s12899-014-0006-0

    Figure Lengend Snippet: Influence of maternal fat supplementation on skeletal muscle enzymatic activity. (A) Enzymatic activities of total lactate dehydrogenase (LDH) and isocitrate dehydrogenase (ICDH) and their ratios in the bicep femoris . (B) Muscular concentrations and ratio of nicotinamide adenine dinucleotide (NADH) as well as its reduced form NAD + on 7-day old offspring exposed to a control (white; n = 7) or high fat (black; n = 7) maternal diet throughout gestation. Bar graphs illustrate means ± SEM (#p

    Article Snippet: NAD+/NADH ratio was determined using a colorimetric commercial kit (Sigma-Aldrich Co LLC, Gillingham, UK) and normalised to the quantity of tissue dissected i.e. moles per gram of protein.

    Techniques: Activity Assay

    (a) Superoxide dismutase, (b) catalase, (c) reduced glutathione, (d) glutathione disulfide, and (e) NAD + / NADH of Escherichia coli ( ATCC 25922), Pseudomonas aeruginosa ( ATCC 27853), and Staphylococcus aureus ( ATCC 29213) exposed to protocatechuic acid (4× MIC ). Values are mean ± SEM of three determinations and are statistically significant at p

    Journal: MicrobiologyOpen

    Article Title: Involvement of oxidative stress in protocatechuic acid‐mediated bacterial lethality, et al. Involvement of oxidative stress in protocatechuic acid‐mediated bacterial lethality

    doi: 10.1002/mbo3.472

    Figure Lengend Snippet: (a) Superoxide dismutase, (b) catalase, (c) reduced glutathione, (d) glutathione disulfide, and (e) NAD + / NADH of Escherichia coli ( ATCC 25922), Pseudomonas aeruginosa ( ATCC 27853), and Staphylococcus aureus ( ATCC 29213) exposed to protocatechuic acid (4× MIC ). Values are mean ± SEM of three determinations and are statistically significant at p

    Article Snippet: 2.6.4 NAD+ /NADH The NAD+ /NADH ratio of bacteria cells was assessed using the Sigma‐Aldrich assay kit (MAK037).

    Techniques:

    Measurement of the in vivo NAD/NADH ratio of S. coelocolor strains with or without light illumination. Intracellular NAD/NADH in illuminated strains was much higher than that in strains cultured in dark. The NAD/NADH concentration in gdhA overexpressed strains was lower 17% than that in the control strains when both of them were cultured in dark. When both of them were cultured in light, the NAD/NADH ratio in the gdhA overexpressed strain was much lower than that in the control strain. n=3, mean±S.D., p

    Journal: Biochemistry and Biophysics Reports

    Article Title: Alpha-ketoglutarate protects Streptomyces coelicolor from visible light-induced phototoxicity

    doi: 10.1016/j.bbrep.2016.11.002

    Figure Lengend Snippet: Measurement of the in vivo NAD/NADH ratio of S. coelocolor strains with or without light illumination. Intracellular NAD/NADH in illuminated strains was much higher than that in strains cultured in dark. The NAD/NADH concentration in gdhA overexpressed strains was lower 17% than that in the control strains when both of them were cultured in dark. When both of them were cultured in light, the NAD/NADH ratio in the gdhA overexpressed strain was much lower than that in the control strain. n=3, mean±S.D., p

    Article Snippet: 2.6 NAD/NADH ratio measurement NAD/NADH ratio was determined by using NAD/NADH Quantification Kit (Sigma-Aldrich, St. Louis, MO, USA).

    Techniques: In Vivo, Cell Culture, Concentration Assay

    Differences in intracellular NAD(H) level between SM-resistant and -sensitive cell lines. The levels of NAD + , NADH, NAD + /NADH ratio and total NAD were determined using NAD/NADH Quantification Kit (Sigma). Therefore, cells were exposed to1.8 μM, 3.6 μM or 7.2 μM SM as well as EtOH as control (0 μM) for 24 hours. Afterwards, the assay was performed according to user’s manual. The results are shown for HaCaT (white) and HaCaT/SM (black) as mean ± SD of triplicates. Significant differences (p

    Journal: Toxicology letters

    Article Title: Sulfur mustard resistant keratinocytes obtained elevated glutathione levels and other changes in the antioxidative defense mechanism

    doi: 10.1016/j.toxlet.2017.11.024

    Figure Lengend Snippet: Differences in intracellular NAD(H) level between SM-resistant and -sensitive cell lines. The levels of NAD + , NADH, NAD + /NADH ratio and total NAD were determined using NAD/NADH Quantification Kit (Sigma). Therefore, cells were exposed to1.8 μM, 3.6 μM or 7.2 μM SM as well as EtOH as control (0 μM) for 24 hours. Afterwards, the assay was performed according to user’s manual. The results are shown for HaCaT (white) and HaCaT/SM (black) as mean ± SD of triplicates. Significant differences (p

    Article Snippet: NAD/NADH determination was performed according to user’s manual using NAD/NADH Quantitation Kit (Sigma).

    Techniques:

    A proposed model for the role of SIRT/ PGC-1/ PPAR alpha network in regulation of radiation-induced cardiac injury. Irradiation impairs the mitochondrial complex I activity resulting in NAD + homeostasis alteration. A change in the level of NAD + /NADH reduces the activity of SIRT3 and enhances the acetylation state of mitochondrial proteins. NAD + /NADH alteration also affects the SIRT1 activity and impairs the PGC-1/ PPAR alpha transcription complex via an increased level of acetylated (inactive) PGC-1. Deactivation of PGC-1/ PPAR alpha is associated with a low level of myocardial metabolism, elevated oxidative damage and accelerated senescence contributing to the radiation-induced cardiac injury.

    Journal: International Journal of Molecular Sciences

    Article Title: Hyperacetylation of Cardiac Mitochondrial Proteins Is Associated with Metabolic Impairment and Sirtuin Downregulation after Chronic Total Body Irradiation of ApoE -/- Mice

    doi: 10.3390/ijms20205239

    Figure Lengend Snippet: A proposed model for the role of SIRT/ PGC-1/ PPAR alpha network in regulation of radiation-induced cardiac injury. Irradiation impairs the mitochondrial complex I activity resulting in NAD + homeostasis alteration. A change in the level of NAD + /NADH reduces the activity of SIRT3 and enhances the acetylation state of mitochondrial proteins. NAD + /NADH alteration also affects the SIRT1 activity and impairs the PGC-1/ PPAR alpha transcription complex via an increased level of acetylated (inactive) PGC-1. Deactivation of PGC-1/ PPAR alpha is associated with a low level of myocardial metabolism, elevated oxidative damage and accelerated senescence contributing to the radiation-induced cardiac injury.

    Article Snippet: NAD+ / NADH Assay Mitochondrial NAD+ /NADH levels were quantified with a commercially available kit (MAK037, Sigma Chemical, St. Louis, Missouri, USA) according to the manufacturer’s instructions.

    Techniques: Pyrolysis Gas Chromatography, Irradiation, Activity Assay

    Analysis of the mitochondrial NAD + , NADH and NAD + /NADH and Acetyl-CoA. The concentration of NAD + , NADH and NAD + /NADH ( A ) and Acetyl-CoA ( B ) was compared in samples from irradiated and control groups. The error bars represent standard error of the mean (± SEM) (t-test; * p

    Journal: International Journal of Molecular Sciences

    Article Title: Hyperacetylation of Cardiac Mitochondrial Proteins Is Associated with Metabolic Impairment and Sirtuin Downregulation after Chronic Total Body Irradiation of ApoE -/- Mice

    doi: 10.3390/ijms20205239

    Figure Lengend Snippet: Analysis of the mitochondrial NAD + , NADH and NAD + /NADH and Acetyl-CoA. The concentration of NAD + , NADH and NAD + /NADH ( A ) and Acetyl-CoA ( B ) was compared in samples from irradiated and control groups. The error bars represent standard error of the mean (± SEM) (t-test; * p

    Article Snippet: NAD+ / NADH Assay Mitochondrial NAD+ /NADH levels were quantified with a commercially available kit (MAK037, Sigma Chemical, St. Louis, Missouri, USA) according to the manufacturer’s instructions.

    Techniques: Concentration Assay, Irradiation, T-Test

    Systemic exposure to 18% oxygen regulates levels of NAD +, NADH, lactate, LDH and SUR2A in the heart. Bar graphs depicting PO 2 (O 2 ), PCO 2 (CO 2 ) and haematocrit (HCT) in the blood as well as ATP (ATP), lactate (Lactate), LDH, total NAD (NADt), NADH (NADH), NAD + (NAD +) and SUR2A (SUR2A) in the myocardial tissue; inset in SUR2A bar graph represents original Western blots for SUR2A under labelled conditions. Each bar is a mean ± SEM (n = 3–5). *P

    Journal: Biochimica et Biophysica Acta

    Article Title: Mild hypoxia in vivo regulates cardioprotective SUR2A: A role for Akt and LDH

    doi: 10.1016/j.bbadis.2015.01.001

    Figure Lengend Snippet: Systemic exposure to 18% oxygen regulates levels of NAD +, NADH, lactate, LDH and SUR2A in the heart. Bar graphs depicting PO 2 (O 2 ), PCO 2 (CO 2 ) and haematocrit (HCT) in the blood as well as ATP (ATP), lactate (Lactate), LDH, total NAD (NADt), NADH (NADH), NAD + (NAD +) and SUR2A (SUR2A) in the myocardial tissue; inset in SUR2A bar graph represents original Western blots for SUR2A under labelled conditions. Each bar is a mean ± SEM (n = 3–5). *P

    Article Snippet: 2.7 Measurement of NAD/NADH NAD/NADH was measured in heart tissue using NAD/NADH kit (Abcam, Cambridge, UK) according to the manufacturer's instruction.

    Techniques: Western Blot

    Metformin inhibits mitochondrial electron transport complex I to limit PM-induced ROS generation from complex III. ( A-C ) A murine alveolar macrophage cell line (MHS) was stably transfected with a lentivirus encoding GFP and NDI1, a yeast protein capable of transferring electrons from NADH to complex II/III but incapable of ROS generation, or GFP alone. These cells were exposed to PM (10 μg/m 3 ) in the presence or absence of metformin and oxygen consumption (Seahorse XF Analyzer) and the oxidation of MitoSOX were measured 4 hours later, and the release of IL-6 into the media was measured 24 hours later (minimum of 8 replicates per measurement, * p

    Journal: Cell metabolism

    Article Title: Metformin Targets Mitochondrial Electron Transport to Reduce Air Pollution-Induced Thrombosis

    doi: 10.1016/j.cmet.2018.09.019

    Figure Lengend Snippet: Metformin inhibits mitochondrial electron transport complex I to limit PM-induced ROS generation from complex III. ( A-C ) A murine alveolar macrophage cell line (MHS) was stably transfected with a lentivirus encoding GFP and NDI1, a yeast protein capable of transferring electrons from NADH to complex II/III but incapable of ROS generation, or GFP alone. These cells were exposed to PM (10 μg/m 3 ) in the presence or absence of metformin and oxygen consumption (Seahorse XF Analyzer) and the oxidation of MitoSOX were measured 4 hours later, and the release of IL-6 into the media was measured 24 hours later (minimum of 8 replicates per measurement, * p

    Article Snippet: In the first method, 100,000 cells were used for measurement of the NAD+ /NADH using an assay kit from Abcam (ab65348) according to the manufacturer’s instructions.

    Techniques: Stable Transfection, Transfection, Transferring

    The effect of systemic exposure to 15% oxygen on different parameters in blood and myocardial tissue. Bar graphs depicting PO 2 (O 2 ), PCO 2 (CO2) and haematocrit (HCT) in the blood as well as ATP (ATP), lactate (Lactate), total NAD (NADt), NADH (NADH) and NAD+ (NAD+) in the myocardial tissue. Each bar is a mean ± S.E.M. ( n = 3–5). * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Exposure to 15% oxygen in vivo up‐regulates cardioprotective SUR2A without affecting ERK1/2 and AKT: a crucial role for AMPK

    doi: 10.1111/jcmm.13064

    Figure Lengend Snippet: The effect of systemic exposure to 15% oxygen on different parameters in blood and myocardial tissue. Bar graphs depicting PO 2 (O 2 ), PCO 2 (CO2) and haematocrit (HCT) in the blood as well as ATP (ATP), lactate (Lactate), total NAD (NADt), NADH (NADH) and NAD+ (NAD+) in the myocardial tissue. Each bar is a mean ± S.E.M. ( n = 3–5). * P

    Article Snippet: Measurement of NAD/NADH in the heart NAD/NADH was measured in heart tissue using NAD/NADH kit (Abcam, Cambridge, UK) according to the manufacturer's instruction.

    Techniques:

    The possible mechanism of the mitochondrial complex I-dependent protective effect of metformin on ATO-induced hepatotoxicity. Metformin increased the intracellular NADH/NAD + ratio by inhibiting mitochondrial respiratory chain complex I, further decreasing the intracellular ROS induced by ATO

    Journal: Cell Death & Disease

    Article Title: Metformin ameliorates arsenic trioxide hepatotoxicity via inhibiting mitochondrial complex I

    doi: 10.1038/cddis.2017.482

    Figure Lengend Snippet: The possible mechanism of the mitochondrial complex I-dependent protective effect of metformin on ATO-induced hepatotoxicity. Metformin increased the intracellular NADH/NAD + ratio by inhibiting mitochondrial respiratory chain complex I, further decreasing the intracellular ROS induced by ATO

    Article Snippet: NADH/NAD+ determination Experiments to determine the NADH/NAD+ ratio in AML12 cells exposed to metformin and/or ATO were carried out using the NAD+ /NADH assay kit from Abcam (ab65348, Cambridge, MA, USA) according to the manufacturer’s instructions.

    Techniques:

    Effects of metformin and ATO on mitochondrial complex I and the NADH/NAD + ratio in AML12 cells. The AML12 cells were pretreated with metformin (5 mM) and/or ATO (6 μ M) for 5 h. Then, the media was replaced by the modified DMEM (Seahorse Bioscience) with pyruvate ( a ) or glutamate ( b ). Rotenone was added to abrogate the function of complex I, and succinate was further added to activate the OCR. The values of the last detection are represented as the means±S.D. (6 replicates for per group). ( c ) The media was replaced by the modified DMEM (Seahorse Bioscience) without any substrate of the mitochondrial respiratory chain. Then, pyruvate and glutamate were added to activate the OCR of AML12 cells. Rotenone was added to abrogate the function of complex I, and succinate was further added to activate the OCR. The values of the fourth ( d ) and last ( e ) detections are represented as the means±S.D. (six replicates for per group). ( f ) The NADH/NAD + ratios in AML12 cells treated with the agents ATO, metformin, rotenone, ATO+metformin and ATO+rotenone or cultured with low glucose DMEM/F12 and/or ATO for 24 h were detected. The values are represented as the means±S.D. (three replicates for per group; * P

    Journal: Cell Death & Disease

    Article Title: Metformin ameliorates arsenic trioxide hepatotoxicity via inhibiting mitochondrial complex I

    doi: 10.1038/cddis.2017.482

    Figure Lengend Snippet: Effects of metformin and ATO on mitochondrial complex I and the NADH/NAD + ratio in AML12 cells. The AML12 cells were pretreated with metformin (5 mM) and/or ATO (6 μ M) for 5 h. Then, the media was replaced by the modified DMEM (Seahorse Bioscience) with pyruvate ( a ) or glutamate ( b ). Rotenone was added to abrogate the function of complex I, and succinate was further added to activate the OCR. The values of the last detection are represented as the means±S.D. (6 replicates for per group). ( c ) The media was replaced by the modified DMEM (Seahorse Bioscience) without any substrate of the mitochondrial respiratory chain. Then, pyruvate and glutamate were added to activate the OCR of AML12 cells. Rotenone was added to abrogate the function of complex I, and succinate was further added to activate the OCR. The values of the fourth ( d ) and last ( e ) detections are represented as the means±S.D. (six replicates for per group). ( f ) The NADH/NAD + ratios in AML12 cells treated with the agents ATO, metformin, rotenone, ATO+metformin and ATO+rotenone or cultured with low glucose DMEM/F12 and/or ATO for 24 h were detected. The values are represented as the means±S.D. (three replicates for per group; * P

    Article Snippet: NADH/NAD+ determination Experiments to determine the NADH/NAD+ ratio in AML12 cells exposed to metformin and/or ATO were carried out using the NAD+ /NADH assay kit from Abcam (ab65348, Cambridge, MA, USA) according to the manufacturer’s instructions.

    Techniques: Modification, Cell Culture

    CR attenuates AngII-induced MMP2 expression through VSMC-SIRT1–dependent H3K9 deacetylation in the Mmp2 promoter. (A, left) Western blots of H3K9 acetylation (H3K9ac), H3, H4K16ac, and H4 in aortas of Apoe −/− mice and SVKO; Apoe −/− mice. (Right) Densitometry was quantified and normalized to the AL-Con group. n = 4 per group. (B and C) ChIP of H3K9ac (B) and H4K16ac (C) on the Mmp2 promoter in Apoe −/− mouse aortas. Four regions were detected: −919 ∼ −784, −669 ∼ −485, −404 ∼ −261, and −260 ∼ −105 bp. n = 4 per group. (D) ChIP assays of H3K9ac on the Mmp2 promoter upon saline (Con) or AngII infusion. H3K9ac in the regions of −919 to −784, −669 to −485, −404 to −261, and −260 to −105 bp of the Mmp2 promoter in the aortas of Apoe −/− and SVKO; Apoe −/− mice is shown. n = 3 per group. (E) ChIP of H4K16ac on the Mmp2 promoter in aortic tissues of AL-Con, CR-Con, AL-AngII, and CR-AngII Apoe −/− mice. n = 3 per group. (F) Western blotting examination of aortic SIRT1 expression upon saline (Con) or AngII infusion for 4 wk in Apoe −/− mice. The quantification of Western blots is provided. n = 3 per group. (G and H) SIRT1 deacetylase activity (G) and NAD + /NADH ratio (H) in the aortas of Apoe −/− mice in the indicated groups were examined. n = 6 per group. (I) SIRT1 enrichment in the Mmp2 promoter was markedly higher than normal IgG in the aortas of AL-Con Apoe −/− mice. n = 3 per group. (J) SIRT1 enrichment in the regions of −919 to −784, −669 to −485, −404 to −261, and −260 to −105 bp of the Mmp2 promoter in the aortas of Apoe −/− mice in the indicated groups. n = 3 per group. Three independent experiments were performed for ChIP assays. All values are shown as the means ± SEM. *, P

    Journal: The Journal of Experimental Medicine

    Article Title: Calorie restriction protects against experimental abdominal aortic aneurysms in mice

    doi: 10.1084/jem.20151794

    Figure Lengend Snippet: CR attenuates AngII-induced MMP2 expression through VSMC-SIRT1–dependent H3K9 deacetylation in the Mmp2 promoter. (A, left) Western blots of H3K9 acetylation (H3K9ac), H3, H4K16ac, and H4 in aortas of Apoe −/− mice and SVKO; Apoe −/− mice. (Right) Densitometry was quantified and normalized to the AL-Con group. n = 4 per group. (B and C) ChIP of H3K9ac (B) and H4K16ac (C) on the Mmp2 promoter in Apoe −/− mouse aortas. Four regions were detected: −919 ∼ −784, −669 ∼ −485, −404 ∼ −261, and −260 ∼ −105 bp. n = 4 per group. (D) ChIP assays of H3K9ac on the Mmp2 promoter upon saline (Con) or AngII infusion. H3K9ac in the regions of −919 to −784, −669 to −485, −404 to −261, and −260 to −105 bp of the Mmp2 promoter in the aortas of Apoe −/− and SVKO; Apoe −/− mice is shown. n = 3 per group. (E) ChIP of H4K16ac on the Mmp2 promoter in aortic tissues of AL-Con, CR-Con, AL-AngII, and CR-AngII Apoe −/− mice. n = 3 per group. (F) Western blotting examination of aortic SIRT1 expression upon saline (Con) or AngII infusion for 4 wk in Apoe −/− mice. The quantification of Western blots is provided. n = 3 per group. (G and H) SIRT1 deacetylase activity (G) and NAD + /NADH ratio (H) in the aortas of Apoe −/− mice in the indicated groups were examined. n = 6 per group. (I) SIRT1 enrichment in the Mmp2 promoter was markedly higher than normal IgG in the aortas of AL-Con Apoe −/− mice. n = 3 per group. (J) SIRT1 enrichment in the regions of −919 to −784, −669 to −485, −404 to −261, and −260 to −105 bp of the Mmp2 promoter in the aortas of Apoe −/− mice in the indicated groups. n = 3 per group. Three independent experiments were performed for ChIP assays. All values are shown as the means ± SEM. *, P

    Article Snippet: NAD+ /NADH level measurement Freshly isolated mouse aortas were used for measuring aortic NAD+ /NADH levels following the manufacturer’s instructions (ab65348; Abcam).

    Techniques: Expressing, Western Blot, Mouse Assay, Chromatin Immunoprecipitation, Histone Deacetylase Assay, Activity Assay

    Intracellular total NAD levels in platelet lysates. Total NAD was measured in platelet lysates during storage of PC. Dotted lines indicate when addition of NR or vehicle solution was performed. Samples were collected on Days 1, 5, 7, 12, 19, and 23 from PC supplemented with NR ( ○ ) or control (●). Data are shown as mean with SD (n = 6), ****p

    Journal: Transfusion

    Article Title: The senotherapeutic nicotinamide riboside raises platelet nicotinamide adenine dinucleotide levels but cannot prevent storage lesion

    doi: 10.1111/trf.15556

    Figure Lengend Snippet: Intracellular total NAD levels in platelet lysates. Total NAD was measured in platelet lysates during storage of PC. Dotted lines indicate when addition of NR or vehicle solution was performed. Samples were collected on Days 1, 5, 7, 12, 19, and 23 from PC supplemented with NR ( ○ ) or control (●). Data are shown as mean with SD (n = 6), ****p

    Article Snippet: Total nicotinamide adenine dinucleotide levels Extraction and detection of total nicotinamide adenine dinucleotide (NAD) levels was performed using an NAD/NADH colorimetric assay kit (Abcam) according to the manual with minor adaptations.

    Techniques:

    Effect of MeHg on the production of NAD(H). Undifferentiated PC12 cells were treated with 0μM (white) or 2μM (black) MeHg for 60 min, and then the amounts of NAD + (A) and NADH (B) were measured by spectrophotometry using an NAD

    Journal: Toxicological Sciences

    Article Title: Methylmercury Impairs Canonical Dopamine Metabolism in Rat Undifferentiated Pheochromocytoma (PC12) Cells by Indirect Inhibition of Aldehyde Dehydrogenase

    doi: 10.1093/toxsci/kfv001

    Figure Lengend Snippet: Effect of MeHg on the production of NAD(H). Undifferentiated PC12 cells were treated with 0μM (white) or 2μM (black) MeHg for 60 min, and then the amounts of NAD + (A) and NADH (B) were measured by spectrophotometry using an NAD

    Article Snippet: NADH/NAD was extracted using 400 µl NADH/NAD Extraction Buffer (Sigma) by 2 freeze/thaw cycles (20 min on dry ice followed by 10 min at room temperature).

    Techniques: Spectrophotometry