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Triphosphopyridine nucleotide serves as an electron carrier in a number of reactions being alternately oxidized NADP and reduced NADPH
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NADP is the oxidized form of the electron donor nicotinamide adenine dinucleotide phosphate NADPH Item No 9000743 It serves as a cofactor in various biological reactions In addition the balance
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Image Search Results

Journal: Redox Biology
Article Title: Cyanidin and delphinidin modulate inflammation and altered redox signaling improving insulin resistance in high fat-fed mice
doi: 10.1016/j.redox.2018.05.012
Figure Lengend Snippet: Effects of supplementation with an AC-rich blend on parameters of liver oxidative stress in HFD mice. Mice were fed a control diet (empty bars), the control diet supplemented with 40 mg AC/kg body weight (dashed bars), a HFD (black bars), or the HFD supplemented with 40 mg AC/kg body weight (red bars). At week 14 on the corresponding diets the following parameters were measured in liver by Western blot: gp91phox, NOX3, NOX4 and HNE-protein adducts, Bands were quantified and values referred to β-actin levels (loading control. Results for CA, HF and HFA40 were referred to control group values (C). Results are shown as mean ± SE of 6 animals/group. Values having different superscripts are significantly different (p
Article Snippet: Antibodies for 4-hydroxynonenal (4-HNE) (ab46545),
Techniques: Mouse Assay, Western Blot

Journal: Antioxidants & Redox Signaling
Article Title: Novel Role of NADPH Oxidase in Angiogenesis and Stem/Progenitor Cell Function
doi: 10.1089/ars.2009.2582
Figure Lengend Snippet: Role of NADPH oxidase in redox signaling linked to neovascularization. NADPH oxidase (Nox) is activated by angiogenesis factors such as VEGF or hypoxia, ischemia, cytokines, and angiotensin II (Ang II). ROS levels are balanced by ROS-generating enzyme, NADPH oxidase, and antioxidant enzymes such as superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPx). Nox-derived ROS activate redox-sensitive kinases/enzymes, including Akt, Src, MAP kinases and eNOS, or inactivate specific protein tyrosine phosphatases (PTPs), which negatively regulate protein tyrosine kinases (PTKs) or protein tyrosine kinases (PTKs) through oxidation of proteins. In addition, ROS activate various redox-sensitive transcription factors. The overall biologic impact of Nox activation thus involves endothelial cell and stem/progenitor cell proliferation, migration, differentiation, senescence, apoptosis, and gene expression, thereby regulating angiogenesis and vasculogenesis, which contribute to neovascularization. NF-κB, nuclear factor kappa B; AP1, activator protein 1; HIF1, hypoxia inducible factor 1; MAPK, mitogen-activated protein kinase; eNOS, endothelial nitric oxide synthase; PTP-SH, reduced form of protein tyrosine phosphatases; PTP-SOH, oxidized form of protein tyrosine phosphatases; RTK, receptor tyrosine kinases; PTK, protein tyrosine kinases.
Article Snippet:
Techniques: Derivative Assay, Activation Assay, Migration, Expressing

Journal: Antioxidants & Redox Signaling
Article Title: Novel Role of NADPH Oxidase in Angiogenesis and Stem/Progenitor Cell Function
doi: 10.1089/ars.2009.2582
Figure Lengend Snippet: Role of NADPH oxidase in ischemia-induced neovascularization. Postnatal new blood vessel formation depends on angiogenesis (formation of new capillaries from preexisting vessels) and vasculogenesis ( de novo new-vessel formation through bone marrow (BM)-derived angiogenic stem/progenitor cells. In response to ischemia, Nox2-based NADPH oxidase expression and reactive oxygen species (ROS) production is increased in BM, which in turn stimulates stem/progenitor cell mobilization from BM to the circulation. NADPH oxidase (NOX)-derived ROS are also involved in stem/progenitor homing and the angiogenic capacity to promote neovascularization of ischemic tissues. Angiogenesis growth factors such as VEGF, produced in response to ischemia, also stimulate endothelial cell (EC) migration, proliferation, and tube formation through an increase in NADPH oxidase–derived ROS to promote angiogenesis. Thus, NADPH oxidase plays an important role in postnatal angiogenesis and stem/progenitor cell function.
Article Snippet:
Techniques: Derivative Assay, Expressing, Produced, Migration, Cell Function Assay

Journal: Antioxidants & Redox Signaling
Article Title: Novel Role of NADPH Oxidase in Angiogenesis and Stem/Progenitor Cell Function
doi: 10.1089/ars.2009.2582
Figure Lengend Snippet: Possible mechanism by which NADPH oxidase is involved in stem/progenitor cell mobilization from bone marrow, and homing and angiogenesis at the site of neovascularization. In response to ischemic stress, hematopoietic and angiogenic factors are increased in bone marrow, which in turn activates NADPH oxidase (NOX) to exit quiescent stem cells from endosteal niche to stimulate proliferation and differentiation. This mechanism may be mediated through activation of matrix metalloproteinase (MMP)-9 and subsequent release of soluble Kit ligand (sKitL), which mobilizes proangiogenic stem/progenitor cells from niches. Furthermore, NOX-derived reactive oxygen species (ROS) may activate redox signaling events involved in proliferation, migration, differentiation, and survival of stem/progenitor cells. As a consequence, stem/progenitor cells are mobilized from bone marrow to the circulation and homed to the ischemic sites through the mechanism, dependent on adhesion and migration. NOX-derived ROS may be involved in expression of adhesion molecules on endothelial surface as well as SDF-1/CXCR4– or IL-8/CXCR2–mediated stem/progenitor cell migration, which may contribute to neovascularization with endothelial cell (EC)-mediated angiogenesis. (VEGF, vascular endothelial growth factor; PlGF, placental growth factor; SDF-1, stroma-derived factor-1; G-CSF, granulocyte colony-stimulating factor; GM-CSF, granulocyte–macrophage colony-stimulating factor; ICAM1, intercellular adhesion molecule 1; VCAM-1, vascular cell adhesion molecule 1).
Article Snippet:
Techniques: Activation Assay, Derivative Assay, Migration, Expressing

Journal: Nature Communications
Article Title: The histone demethylase Phf2 acts as a molecular checkpoint to prevent NAFLD progression during obesity
doi: 10.1038/s41467-018-04361-y
Figure Lengend Snippet: Phf2 and ChREBP decrease oxidative stress through Nrf2 activation. Mice, injected with either GFP or Phf2 overexpressing adenovirus, were studied 3 weeks later in fed state. a Representative Western blot analysis of Nrf2 protein content and Nrf2-regulated genes ( n = 10 mice per group). b – d Nrf2 was inhibited in cultured hepatocytes overexpressing Phf2. b Western blot analysis of proteins involved in oxidative stress defenses ( n = 3). GSH and NADPH contents ( c ) in addition to ROS levels and hepatocyte apoptosis ( d ) were determined after incubation with 480 μM palmitate for 24 h ( n = 3). e UCSC genome browser image illustrating normalized tag counts for Phf2, ChREBP, H3K9me2, H3K4me3, and RNA polII at the Nrf2 promoter. ( f , g ) Phf2 was inhibited in cultured hepatocytes. ChIP for H3K9me2, ChREBP, and RNA polII, in addition to chromatin accessibility at the Nrf2 promoter and Nrf2 activity on the ARE-luc construct shown ( n = 3). h – j ChREBP expression was inhibited in cultured hepatocytes overexpressing Phf2. h Representative western blot analysis showing the contribution of ChREBP to the regulation of Nrf2 and Nrf2-regulated gene expression. i Nrf2 activity on the ARE-luc construct shown. j Relative ROS levels and measurement of hepatocyte apoptosis were determined after incubation with 480 μM palmitate for 24 h ( n = 3). All error bars represent mean ± SEM. Statistical analyses were made using Anova, followed by Bonferonni’s test. * P
Article Snippet: The NADP/NADPH ratio was determined using the
Techniques: Activation Assay, Mouse Assay, Injection, Western Blot, Cell Culture, Incubation, Chromatin Immunoprecipitation, Activity Assay, Construct, Expressing

Journal: Nature Communications
Article Title: The histone demethylase Phf2 acts as a molecular checkpoint to prevent NAFLD progression during obesity
doi: 10.1038/s41467-018-04361-y
Figure Lengend Snippet: Phf2-driven Nrf2 activation protects liver from fibrogenesis. GFP or Phf2 were overexpressed in the liver of Wild Type (WT) and Nrf2 knockout (Nrf2-KO) mice. Mice were studied 3 weeks later in the fed state. a Western blot analysis of liver extracts from WT or Nrf2-KO mice ( n = 10 mice per group). b (Left) Oral glucose tolerance test and insulin levels during the OGTT test. (right) Insulin tolerance test ( n = 10 per group). c Expression of coll-Ia1, α-SMA, and TIMP-1 ( n = 10 mice per group). d Percentage of fibrotic area and percentage of apoptotic hepatocytes ( n = 10 per group). e NADPH and GSH contents ( n = 10 mice per group). f In vivo bioluminescent response of the PCL-2 probe to H 2 O 2 . Representative image for mice injected with the PCL-2 probe is shown ( n = 6 mice per group). g Levels of carbonylated proteins ( n = 10 mice per group). All error bars represent mean ± SEM. Statistical analyses were made using unpaired t -test ( b ) or Anova, followed by Bonferonni’s test ( c – e, g ). * P
Article Snippet: The NADP/NADPH ratio was determined using the
Techniques: Activation Assay, Knock-Out, Mouse Assay, Western Blot, Expressing, In Vivo, Injection

Journal: Nature Communications
Article Title: The histone demethylase Phf2 acts as a molecular checkpoint to prevent NAFLD progression during obesity
doi: 10.1038/s41467-018-04361-y
Figure Lengend Snippet: Phf2 diverts glucose fluxes to protect liver from oxidative stress. Mice, injected with either GFP or Phf2 overexpressing adenovirus, were studied 3 weeks later in the fed state. a Glucose and oleate oxidation rate determined by measuring the production of 14 CO 2 from 14 C-glucose or 14 C-oleate for 4 h ( n = 8 mice per group). b Basal glutamate/malate and succinate-driven mitochondrial respiration, which provide respectively electrons to the complex I and II of the mitochondrial reaction chain ( n = 6 mice per group). c Relative ROS and carbonylated protein levels ( n = 12 mice per group). d Metabolomic KEGG pathway enrichment analysis ( n = 15 mice per group). e Heat map of metabolic intermediates of the PPP, serine, glycine, and GSH biosynthetic pathways ( n = 15 mice per group). f Western blot analysis of proteins involved in oxidative stress defenses ( n = 12 mice per group). g Liver NADPH content ( n = 10 mice per group). h Measurement of Gpx activity ( n = 10 mice per group). i Expression of genes involved in GSH synthesis ( n = 10 mice per group). All error bars represent mean ± SEM. Statistical analyses were made using unpaired t -test. * P
Article Snippet: The NADP/NADPH ratio was determined using the
Techniques: Mouse Assay, Injection, Western Blot, Activity Assay, Expressing

Journal: International Journal of Oncology
Article Title: ABT737 reverses cisplatin resistance by targeting glucose metabolism of human ovarian cancer cells
doi: 10.3892/ijo.2018.4476
Figure Lengend Snippet: Redox homeostasis in SKOV3/DDP cells is maintained intrinsically by pairing oxidative phosphorylation with pentose phosphate pathway. (A) Cellular NADPH content and (B) NADPH/NADP + ratio, (C) GSH and (D) GSSG contents, (E) total GSH (GSH plus GSSG) level and (F) GSH/GSSG ratio determined using enzymatic assays. (G) The expression level G6PD gene was detected using reverse transcription-quantitative polymerase chain reaction. (H) G6PD protein expression level was determined using western blotting, and the enzymatic activity was analyzed using a G6PD assay kit. The data are representative of three experiments. (I) Cell viability of SKOV3 or SKOV3/DDP cells was determined using a MTT assay in the presence of G6PD inhibitors (20 μ M 6-AN or 250 μ M DHEA) with or without cisplatin for 24 h. * P
Article Snippet: Then, the cells were subjected to analysis with the
Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Activity Assay, G6PD Assay, MTT Assay

Journal: Experimental Neurobiology
Article Title: The Unreliability of MTT Assay in the Cytotoxic Test of Primary Cultured Glioblastoma Cells
doi: 10.5607/en.2015.24.3.235
Figure Lengend Snippet: Ethanol increases the intracellular NADH concentration. The ratio of NAD/NADH was determined by the NADP+/NADPH Quantification Kit (BioVision, Inc. K347-100, SF, USA) in GBL-13, GBL-15, U87MG and U373MG. This experiment was conducted in four types of cells that were treated with ethanol for 24 h. Values are expressed as mean and SD. * p
Article Snippet: Intracellular nucleotides such as NAD and NADH ratios were detected by the
Techniques: Concentration Assay

Journal: Nature
Article Title: The metabolic function of cyclin D3-CDK6 kinase in cancer cell survival
doi: 10.1038/nature22797
Figure Lengend Snippet: Metabolic changes in T-ALL cells upon inhibition of cyclin D3-CDK6 Flow of glucose-derived carbon into PPP ( a ), and serine pathway ( b ) following D3-CDK6 inhibition in T-ALL KOPTK1 cells expressing wild-type PFKP and PKM2 (KOPTK1-WT), or PFKP-S679E and PKM2-S37E mutants (KOPTK1-EE). Levels of NADPH ( c ), GSH ( d ), ROS ( e ) in T-ALL cell lines upon palbociclib-treatment. f , Apoptosis of cells treated with palbociclib and NAC. g , Apoptosis in KOPTK1-WT and KOPTK1-EE cells upon palbociclib-treatment, or following knockdown of CDK6 ( h ). i , j , In vivo apoptosis of T-ALL cells (in peripheral blood and bone marrow, gated on human CD45 + cells) in mice xenografted with KOPTK1-WT or KOPTK1-EE cells. Data are mean ±s.d. * P
Article Snippet: Intracellular NADPH was measured using
Techniques: Inhibition, Flow Cytometry, Derivative Assay, Expressing, In Vivo, Mouse Assay

Journal: Nature
Article Title: The metabolic function of cyclin D3-CDK6 kinase in cancer cell survival
doi: 10.1038/nature22797
Figure Lengend Snippet: Inhibition of PPP and serine pathway in T-ALL cells a–d , KOPTK1 cells were transduced with vectors encoding shRNA against phosphoserine aminotransferase, the key enzyme in the serine pathway (shPSAT +), or control shRNA (−), and cultured in the presence (+) or absence (−) of dehydroepiandrosterone (DHEA, an inhibitor of pentose phosphate pathway), and the levels of NADPH ( a ), reduced glutathione (GSH, b ), ROS ( c ), and apoptosis ( d , Annexin V staining followed by FACS) were determined. e–h , Similar analysis of MOLT4 cells. i–l , Similar analysis for DND41 cells. m , Immunoblot analysis to gauge the efficiency of PSAT1 knockdown. Tubulin served as a loading control. n–p , The indicated T-ALL cell lines were treated with vehicle (con) or with PPP inhibitors 6-aminonicotinamide (6-AN) or DHEA, and the levels of NADPH ( n ) and ROS ( o ) were assayed. Apoptosis was quantified by Annexin V staining followed by FACS ( p ). a–l , n–p , n=3 biological replicates. Bars, mean values; error bars, s.d. * P
Article Snippet: Intracellular NADPH was measured using
Techniques: Inhibition, Transduction, shRNA, Cell Culture, Staining, FACS

Journal: Cell Death & Disease
Article Title: A new inhibitor of glucose-6-phosphate dehydrogenase blocks pentose phosphate pathway and suppresses malignant proliferation and metastasis in vivo
doi: 10.1038/s41419-018-0635-5
Figure Lengend Snippet: Polydatin Inhibits G6PD causing redox imbalance, which leads to ER stress and cell death. a Viability assay (MTT) of UMSCC103 treated with polydatin (20 µM) and 4μ8c (IRE inhibitor), GSK2606414 (PERK inhibitor) or with siRNA for either IRE1 or PERK, 24 or 48 h posttreatment, respectively. b : menadione is used as positive control; N -acetylcysteine (NAC) is used as ROS scavenger; quantitative assay). c IF with ER-Tracker on UMSCC103 treated with polydatin in combination with NAC, 24 h posttreatment. d G6PD enzymatic assay on UMSCC103 cell lysates (the same assay performed with purified enzyme is found in Fig. S2C). e NADP + /NADPH ratio on polydatin-treated cells. f Invasion assay of UMSCC103 after polydatin treatment. * p
Article Snippet: NADP+ /NADPH ratios in cell lines treated with PD were measured according to the protocol of
Techniques: Viability Assay, MTT Assay, Positive Control, Enzymatic Assay, Purification, Invasion Assay
![MJ25 is an inhibitor of thioredoxin reductase 1 (TrxR1) a. The capability of MJ25 and auranofin to inhibit recombinant, rat-derived TrxR1 in vitro was measured by an NADPH dependent 5,5′-dithiobis-[2-nitrobenzoic acid] (DTNB) assay. b. ARN8 cells were treated with MJ25, auranofin or DMSO, respectively, for the indicated periods of time. TrxR1 inhibition was subsequently assessed in cell lysates by an NADPH and Trx dependent insulin reduction endpoint assay, measuring thiol formation using DTNB. Ratios between MJ25 and DMSO as well as auranofin and DMSO were determined for each point in time. (c and d) ARN8 cells were treated with c. MJ25 or d. auranofin, while in each half of the samples growth media were supplemented with sodium selenite [75 nM] three days prior to seeding as well as during seeding and treatment for 72 hours. Cell viability and clonogenic capacity were determined. e. ROS levels were determined in ARN8 cells 3 hours after the indicated treatment by measuring fluorescence of 2′,7′-dichlorofluorescein (DCF). f. Induction of anti-oxidative proteins by MJ25 and auranofin was investigated in ARN8 cells at the indicated points in time by Western blotting. DMSO served as vehicle control (0 μM). g. ARN8 cells were pre-treated with L-buthionine sulfoximine (BSO) or vehicle (H 2 O) for 72 hours, upon which cells were re-plated in BSO- and vehicle-free growth medium. Cell viability was assessed by SRB assay after 72 hours in the presence of vehicle (DMSO), MJ25 (left panel) or auranofin (middle panel), respectively. Intracellular glutathione (GSH) levels were determined 72 hours after BSO / vehicle treatment (right panel). h. Inhibition of yeast-derived glutathione reductase by MJ25 and auranofin was determined in vitro by measurement of glutathione disulfide (GSSG) reduction.](https://storage.googleapis.com/bioz_article_images/PMC4599284/oncotarget-06-16488-g004.jpg)
Journal: Oncotarget
Article Title: Redox effects and cytotoxic profiles of MJ25 and auranofin towards malignant melanoma cells
doi:
Figure Lengend Snippet: MJ25 is an inhibitor of thioredoxin reductase 1 (TrxR1) a. The capability of MJ25 and auranofin to inhibit recombinant, rat-derived TrxR1 in vitro was measured by an NADPH dependent 5,5′-dithiobis-[2-nitrobenzoic acid] (DTNB) assay. b. ARN8 cells were treated with MJ25, auranofin or DMSO, respectively, for the indicated periods of time. TrxR1 inhibition was subsequently assessed in cell lysates by an NADPH and Trx dependent insulin reduction endpoint assay, measuring thiol formation using DTNB. Ratios between MJ25 and DMSO as well as auranofin and DMSO were determined for each point in time. (c and d) ARN8 cells were treated with c. MJ25 or d. auranofin, while in each half of the samples growth media were supplemented with sodium selenite [75 nM] three days prior to seeding as well as during seeding and treatment for 72 hours. Cell viability and clonogenic capacity were determined. e. ROS levels were determined in ARN8 cells 3 hours after the indicated treatment by measuring fluorescence of 2′,7′-dichlorofluorescein (DCF). f. Induction of anti-oxidative proteins by MJ25 and auranofin was investigated in ARN8 cells at the indicated points in time by Western blotting. DMSO served as vehicle control (0 μM). g. ARN8 cells were pre-treated with L-buthionine sulfoximine (BSO) or vehicle (H 2 O) for 72 hours, upon which cells were re-plated in BSO- and vehicle-free growth medium. Cell viability was assessed by SRB assay after 72 hours in the presence of vehicle (DMSO), MJ25 (left panel) or auranofin (middle panel), respectively. Intracellular glutathione (GSH) levels were determined 72 hours after BSO / vehicle treatment (right panel). h. Inhibition of yeast-derived glutathione reductase by MJ25 and auranofin was determined in vitro by measurement of glutathione disulfide (GSSG) reduction.
Article Snippet: Reduced glutathione (GSH; no. A2084.0005), oxidized glutathione (GSSG; no. G4376), and
Techniques: Recombinant, Derivative Assay, In Vitro, DTNB Assay, Inhibition, End Point Assay, Fluorescence, Western Blot, Sulforhodamine B Assay

Journal: Biomolecules & Therapeutics
Article Title: Niacinamide Protects Skin Cells from Oxidative Stress Induced by Particulate Matter
doi: 10.4062/biomolther.2019.061
Figure Lengend Snippet: Schematic diagram showing the protective action of NIA on PM 2.5 -induced cell damage. NIA protected keratinocytes by suppressing ROS generation by decreasing the NADP/NADPH ratio. Further, NIA prevented oxidative stress-induced molecules damage, including lipid peroxidation, protein carbonylation, and DNA modification. NIA could also stabilize mitochondrial membrane potential by balancing calcium levels, which was disrupted by PM 2.5 . Finally, NIA protected cells from PM 2.5 -induced apoptosis.
Article Snippet: NADP/NADPH assay To determine the ratio of intracellular NADP and
Techniques: Modification

Journal: Biomolecules & Therapeutics
Article Title: Niacinamide Protects Skin Cells from Oxidative Stress Induced by Particulate Matter
doi: 10.4062/biomolther.2019.061
Figure Lengend Snippet: NIA cleared ROS by inhibiting NOX activity induced by PM 2.5 . (A) The ratio of intracellular NADP and NADPH was assessed using NADP/NADPH assay kit. (B) Intracellular ROS was detected after staining of cells with DCF-DA dye. (C) Superoxide generation was detected after dying cells with DHE. NIA diminished superoxide levels induced by PM 2.5 . * p
Article Snippet: NADP/NADPH assay To determine the ratio of intracellular NADP and
Techniques: Activity Assay, Staining

Journal: Mucosal Immunology
Article Title: Cell-mediated reduction of human ?-defensin 1: a major role for mucosal thioredoxin
doi: 10.1038/mi.2013.17
Figure Lengend Snippet: Blockade of the TRX system by auranofin. ( a ) Oxidized hBD-1 was incubated with human TRX, rat thioredoxin reductase, and NADPH while 0 μ M (left) or 2.5 μ M auranofin (right) was added. ( b ) Oxidized hBD-1 was incubated with 5 m M glutathione and 3 μ M GRX without (left) and with 2.5 μ M auranofin (right). Reaction mixtures were analyzed by RP-HPLC as in Figure 1 . GRX, glutaredoxin-1; NADPH, nicotinamide adenine dinucleotide phosphate; oxhBD-1, oxidized human β-defensin 1; redhBD-1, reduced human β-defensin 1; RP-HPLC, reversed-phase, high-performance liquid chromatography; TRX, thioredoxin-1.
Article Snippet: A reaction mixture of 10 μM oxhBD-1 (Peptide Institute, Osaka, Japan), 0.8 mM
Techniques: Incubation, High Performance Liquid Chromatography

Journal: Mucosal Immunology
Article Title: Cell-mediated reduction of human ?-defensin 1: a major role for mucosal thioredoxin
doi: 10.1038/mi.2013.17
Figure Lengend Snippet: Reduction of hBD-1 by different redox enzymes. ( a ) A total of 10 μ M oxhBD-1 was incubated with rat thioredoxin reductase and NADPH. Conversion of oxidized into reduced hBD-1 was monitored by RP-HPLC analysis. Left panel: control; middle panel: addition of 3 μ M TRX; right panel: addition of 3 μ M PDI. ( b ) oxhBD-1 was incubated with 5 m M glutathione (left panel); middle panel: addition of 3 μ M GRX; right panel: addition of 4 μ M MIF. ( c ) oxhBD-1 was incubated with 3 μ M GRX as described for b and incubated for 30, 60, or 90 min. ( d ) oxhBD-1 was incubated with 0.5, 1.5, or 6.0 μ M GRX as described for b and incubated for 30 min. Quantification of hBD-1 forms in c and d was performed by integration of areas under the respective peaks from at least two independent experiments; data are presented as means±s.e.m. GRX, glutaredoxin-1; MIF, macrophage migration inhibitory factor; NADPH, nicotinamide adenine dinucleotide phosphate; oxhBD-1, oxidized human β-defensin 1, PDI, protein disulfide isomerase A1; redhBD-1, reduced human β-defensin 1; RP-HPLC, reversed-phase, high-performance liquid chromatography; TRX, thioredoxin-1. Gray shadow in c indicates that this experiment is quantification of b , middle panel.
Article Snippet: A reaction mixture of 10 μM oxhBD-1 (Peptide Institute, Osaka, Japan), 0.8 mM
Techniques: Incubation, High Performance Liquid Chromatography, Migration

Journal: Applied and Environmental Microbiology
Article Title: Deglycosylation of the Isoflavone C-Glucoside Puerarin by a Combination of Two Recombinant Bacterial Enzymes and 3-Oxo-Glucose
doi: 10.1128/AEM.00607-20
Figure Lengend Snippet: HPLC analysis of DgpA-bound NAD(H). Purified DgpA was denatured with cold methanol, and the dissociated cofactors were analyzed by HPLC. (a) Authentic mixture of cofactors NAD + , NADH, NADP + , and NADPH; (b) DgpA-bound cofactors.
Article Snippet: NAD+ , NADH, NADP+, and
Techniques: High Performance Liquid Chromatography, Purification

Journal: PLoS ONE
Article Title: Oxidative stress caused by activation of NADPH oxidase 4 promotes contrast-induced acute kidney injury
doi: 10.1371/journal.pone.0191034
Figure Lengend Snippet: Effects of Nox4 on iohexol-induced ROS generation. Cells were exposed to iohexol for 0, 1, or 2 h with and without Nox4 knockdown and GKT137831. ( A ) Confocal microscopic images of cells subjected to DHE staining. Original magnification, ×200; scale bar, 20 μm. ( B ) H 2 O 2 , a product of Nox4, was measured by Amplex red assay. The data are the mean ± SD. ** p
Article Snippet: Antibodies used for immunoblotting were as follows:
Techniques: Staining, Amplex Red Assay

Journal: PLoS ONE
Article Title: Oxidative stress caused by activation of NADPH oxidase 4 promotes contrast-induced acute kidney injury
doi: 10.1371/journal.pone.0191034
Figure Lengend Snippet: Proposed model of radiocontrast medium-induced acute kidney injury enhanced by oxidative stress caused by activation of Nox4 in HK-2 cells. Radiocontrast medium induces proximal tubular cell apoptosis through increased oxidative stress by Nox4 activation. Inhibition of Nox4 by genetic knockdown of Nox4 or by treatment with GKT137831 (specific Nox1/4 inhibitor) attenuated radiocontrast-induced proximal tubular cell apoptosis through reduction of MAPK (especially p38 MAPK subfamily) phosphorylation and Bax levels.
Article Snippet: Antibodies used for immunoblotting were as follows:
Techniques: Activation Assay, Inhibition

Journal: PLoS ONE
Article Title: Oxidative stress caused by activation of NADPH oxidase 4 promotes contrast-induced acute kidney injury
doi: 10.1371/journal.pone.0191034
Figure Lengend Snippet: Western blot analysis showing intracellular Nox4 signaling in iohexol-induced apoptosis in HK-2 cells. HK-2 cells were incubated with iohexol for the indicated times (0 min, 5 min, 30 min, 1 h, 2 h, or 3 h). For examination of the long-term viability of HK-2 cells after iohexol exposure, the medium containing iohexol was removed after 5 min, 0.5 h, 1 h, 2 h, or 3 h and then replaced with fresh serum-free medium for 24 h, 23.5 h, 23 h, 22 h, or 21 h (5 min/24 h, 0.5/23.5 h, 1/23 h, 2/22 h or 3/21 h). ( A ) Effects of Nox4 on phosphorylation of MAPK family members (p38, JNK, and ERK), bax, bcl-2, and p65. ( B and C ) Effects of Nox4 knockdown and GKT137831 pretreatment on iohexol-induced apoptosis.
Article Snippet: Antibodies used for immunoblotting were as follows:
Techniques: Western Blot, Incubation

Journal: PLoS ONE
Article Title: Oxidative stress caused by activation of NADPH oxidase 4 promotes contrast-induced acute kidney injury
doi: 10.1371/journal.pone.0191034
Figure Lengend Snippet: Effects of Nox4 knockdown and GKT137831 treatment on iohexol-induced apoptosis and cell death in HK-2 cells. HK-2 cells were cultured to 70–80% confluence, and iohexol was added. Cells were incubated with iohexol at the indicated concentrations for 30 min. For examination of the long-term viability of HK-2 cells after iohexol exposure, medium containing iohexol was removed after 2 h and replaced with fresh serum-free medium for 22 h (2/22 h). ( A ) The apoptotic response of HK-2 cells was assayed by measuring caspase 3/7 activity. ( B ) The viability of HK-2 cells was assayed using ATPlite and MTT assays. The data are the mean ± SD (n = 5). ** p
Article Snippet: Antibodies used for immunoblotting were as follows:
Techniques: Cell Culture, Incubation, Activity Assay, MTT Assay

Journal: PLoS ONE
Article Title: Oxidative stress caused by activation of NADPH oxidase 4 promotes contrast-induced acute kidney injury
doi: 10.1371/journal.pone.0191034
Figure Lengend Snippet: Iohexol-induced differential expression of Nox homologues and time-dependent Nox4 expression in HK-2 cells. (A) HK-2 cells were exposed to iohexol for 30 min. Nox1 , Nox2 , Nox3 , Nox4 , and Nox5 mRNA levels after iohexol exposure were measured by quantitative real-time PCR. ( B) HK-2 cells were exposed to iohexol for 0 min, 1 min, 5 min, 10 min, 30 min, 1 h, 2 h, and 24 h. For examination of the long-term viability of HK-2 cells after iohexol exposure, the medium containing iohexol was removed after 1 or 2 h and replaced with fresh serum-free medium for 23 and 22 h (1/23 and 2/22 h, respectively). Nox4 ( NOX4 ) mRNA expression after iohexol exposure was measured by quantitative real-time PCR. Nox4 protein levels were measured by western blotting. Data are the mean ± SD (n = 5). * p
Article Snippet: Antibodies used for immunoblotting were as follows:
Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot