anti-nqo1 Santa Cruz Biotechnology Search Results


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  • 99
    Santa Cruz Biotechnology nqo1
    BL153 upregulated renal PGC-1 α and <t>NQO1</t> expression. PGC-1 α and NQO1 expression was detected by western blot assay. n = 5; * P
    Nqo1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 99/100, based on 884 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology anti nqo1 a180
    Reduced pyridine nucleotides and dicumarol induce a conformational change in <t>NQO1.</t> (A) Comparison of the ability of antibodies which bind to helix 7 <t>(A180)</t> and antibodies which bind to the C-terminal domain (C-Term) to immunoprecipitate rhNQO1 in the absence and presence of NADH. (B, C) Immunoprecipitation of rhNQO1 with the A180 antibody in the absence and presence of NADH or NADPH. (D) Immunoprecipitation of rhNQO1 by C-Term and A180 antibodies in the absence and presence of dicumarol. (E) The effect of NADH and dicumarol on the migration of rhNQO1 in non-denaturing PAGE. Reaction conditions for immunoprecipitation studies and non-denaturing PAGE are described in Materials and methods .
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    Santa Cruz Biotechnology goat anti nqo1
    Curcumin inhibits <t>NQO1</t> enzymatic activity in vitro and in cells. Enzymatic activity of recombinant NQO1 was determined in the absence (–) or presence (+) of BSA without (–) or with the indicated concentrations of curcumin ( A ) or dicoumarol
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    Santa Cruz Biotechnology rabbit anti nqo1
    Effects of ginsenoside Rd on HO-1 and <t>NQO1</t> expression in rats subjected to myocardial I/R. The representative images of HO-1 and NQO1 are shown. The results are expressed as the mean ± S.E.M. (n = 8 in each group). #P
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    Santa Cruz Biotechnology anti nqo1 sc 271116 antibodies
    Effects of ginsenoside Rd on HO-1 and <t>NQO1</t> expression in rats subjected to myocardial I/R. The representative images of HO-1 and NQO1 are shown. The results are expressed as the mean ± S.E.M. (n = 8 in each group). #P
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    Santa Cruz Biotechnology mouse anti nqo1
    <t>NQO1</t> inhibits PGC-1α proteasomal degradation and increases its protein half-life. (A) HEK-293T cells and HEK-293T cells stably expressing the Flag-tagged β4 (PSMB2) ring subunit were transiently transfected with untagged PGC-1α-
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    Santa Cruz Biotechnology polyclonal goat anti nqo1
    <t>NQO1</t> inhibits PGC-1α proteasomal degradation and increases its protein half-life. (A) HEK-293T cells and HEK-293T cells stably expressing the Flag-tagged β4 (PSMB2) ring subunit were transiently transfected with untagged PGC-1α-
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    Santa Cruz Biotechnology polyclonal anti nqo1
    <t>NQO1</t> inhibits PGC-1α proteasomal degradation and increases its protein half-life. (A) HEK-293T cells and HEK-293T cells stably expressing the Flag-tagged β4 (PSMB2) ring subunit were transiently transfected with untagged PGC-1α-
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    Santa Cruz Biotechnology goat anti nqo1 c19 r20
    <t>NQO1</t> inhibits PGC-1α proteasomal degradation and increases its protein half-life. (A) HEK-293T cells and HEK-293T cells stably expressing the Flag-tagged β4 (PSMB2) ring subunit were transiently transfected with untagged PGC-1α-
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    Santa Cruz Biotechnology goat anti nqo1 antibodies c 19
    <t>NQO1</t> inhibits PGC-1α proteasomal degradation and increases its protein half-life. (A) HEK-293T cells and HEK-293T cells stably expressing the Flag-tagged β4 (PSMB2) ring subunit were transiently transfected with untagged PGC-1α-
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    Santa Cruz Biotechnology mouse anti nqo1 sc 376023 antibodies
    <t>NQO1</t> inhibits PGC-1α proteasomal degradation and increases its protein half-life. (A) HEK-293T cells and HEK-293T cells stably expressing the Flag-tagged β4 (PSMB2) ring subunit were transiently transfected with untagged PGC-1α-
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    Santa Cruz Biotechnology western blot anti nqo1
    3g can elevate the protein level of Nrf2 and <t>NQO1.</t>
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    Santa Cruz Biotechnology nqo1 primary antibodies
    BR specifically inhibits the Nrf2 signaling pathway. A375 cells were treated with BR for 24 hours. (a) Cell survival was assessed using an MTS assay. (b and c) Cells were treated with various concentrations of BR for 24 hours ((b), right; (c) right) or with 50 nM BR for different time interval as indicated ((b), left; (c) left). Total cell extracts were prepared and subjected to Western blot using antibodies for Nrf2, HO-1, <t>NQO1,</t> GSTP1, I κ B α , COX2, NF- κ B, and actin. Data in (a) are shown as mean ± SD; ∗ P
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    Santa Cruz Biotechnology mouse monoclonal anti nqo1 a180
    BR specifically inhibits the Nrf2 signaling pathway. A375 cells were treated with BR for 24 hours. (a) Cell survival was assessed using an MTS assay. (b and c) Cells were treated with various concentrations of BR for 24 hours ((b), right; (c) right) or with 50 nM BR for different time interval as indicated ((b), left; (c) left). Total cell extracts were prepared and subjected to Western blot using antibodies for Nrf2, HO-1, <t>NQO1,</t> GSTP1, I κ B α , COX2, NF- κ B, and actin. Data in (a) are shown as mean ± SD; ∗ P
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    Santa Cruz Biotechnology 200 anti nqo1 mouse monoclonal
    BR specifically inhibits the Nrf2 signaling pathway. A375 cells were treated with BR for 24 hours. (a) Cell survival was assessed using an MTS assay. (b and c) Cells were treated with various concentrations of BR for 24 hours ((b), right; (c) right) or with 50 nM BR for different time interval as indicated ((b), left; (c) left). Total cell extracts were prepared and subjected to Western blot using antibodies for Nrf2, HO-1, <t>NQO1,</t> GSTP1, I κ B α , COX2, NF- κ B, and actin. Data in (a) are shown as mean ± SD; ∗ P
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    Santa Cruz Biotechnology western blot anti nqo1 sc 271116
    BR specifically inhibits the Nrf2 signaling pathway. A375 cells were treated with BR for 24 hours. (a) Cell survival was assessed using an MTS assay. (b and c) Cells were treated with various concentrations of BR for 24 hours ((b), right; (c) right) or with 50 nM BR for different time interval as indicated ((b), left; (c) left). Total cell extracts were prepared and subjected to Western blot using antibodies for Nrf2, HO-1, <t>NQO1,</t> GSTP1, I κ B α , COX2, NF- κ B, and actin. Data in (a) are shown as mean ± SD; ∗ P
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    Santa Cruz Biotechnology nqo1 a180 sc 32793 antibodies
    BR specifically inhibits the Nrf2 signaling pathway. A375 cells were treated with BR for 24 hours. (a) Cell survival was assessed using an MTS assay. (b and c) Cells were treated with various concentrations of BR for 24 hours ((b), right; (c) right) or with 50 nM BR for different time interval as indicated ((b), left; (c) left). Total cell extracts were prepared and subjected to Western blot using antibodies for Nrf2, HO-1, <t>NQO1,</t> GSTP1, I κ B α , COX2, NF- κ B, and actin. Data in (a) are shown as mean ± SD; ∗ P
    Nqo1 A180 Sc 32793 Antibodies, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 99/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology monoclonal nqo1 antibody
    Loss of <t>NQO1</t> expression inhibits cell proliferation and in vivo tumor growth In A and B , A549 and H292 cells were assayed for proliferation rates at 0, 24, 48 and 72 h using the CyQuant cell proliferation kit (Life Technologies). In C , A549 shNQO1 cells (open symbols) and A549 shCtr-R (closed symbols) were subcutaneously injected into flanks of athymic mice at varying concentrations ((1.0 black), (2.5 blue) or (5.0 red) × 10 6 ) cells. Tumor growth was assessed bi/weekly using calipers. In D , A549 shNQO1 and A549 shCtr-R cells were injected subcutaneously into flanks of athymic mice and tumors were measured bi weekly by caliper measurements until a volume of 1000 mm 3 was reached. Kaplan-Meir survival analysis was conducted using GraphPad Prism 6 software. In E , representative photomicrograph of mice in C where mice were injected on the Left flank (L) with A549 shNQO1 cells or on the right flank (R) with A549 shCtr-R cells at the indication concentration of cells. Shown are mice whose tumors were photographed after 32 days. In F , Western Blot for tumor-NQO1 expression and PARP-1 cleavage. Samples were harvested in PARP-lysis buffer as described in “Materials and Methods”. In A , p values for A549 shNQO1 vs A549 shCtr-R cells at 24 h ( p = 0.0031), 48 h ( p
    Monoclonal Nqo1 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology polyclonal goat antibodies against nad
    Loss of <t>NQO1</t> expression inhibits cell proliferation and in vivo tumor growth In A and B , A549 and H292 cells were assayed for proliferation rates at 0, 24, 48 and 72 h using the CyQuant cell proliferation kit (Life Technologies). In C , A549 shNQO1 cells (open symbols) and A549 shCtr-R (closed symbols) were subcutaneously injected into flanks of athymic mice at varying concentrations ((1.0 black), (2.5 blue) or (5.0 red) × 10 6 ) cells. Tumor growth was assessed bi/weekly using calipers. In D , A549 shNQO1 and A549 shCtr-R cells were injected subcutaneously into flanks of athymic mice and tumors were measured bi weekly by caliper measurements until a volume of 1000 mm 3 was reached. Kaplan-Meir survival analysis was conducted using GraphPad Prism 6 software. In E , representative photomicrograph of mice in C where mice were injected on the Left flank (L) with A549 shNQO1 cells or on the right flank (R) with A549 shCtr-R cells at the indication concentration of cells. Shown are mice whose tumors were photographed after 32 days. In F , Western Blot for tumor-NQO1 expression and PARP-1 cleavage. Samples were harvested in PARP-lysis buffer as described in “Materials and Methods”. In A , p values for A549 shNQO1 vs A549 shCtr-R cells at 24 h ( p = 0.0031), 48 h ( p
    Polyclonal Goat Antibodies Against Nad, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 81/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    82
    Santa Cruz Biotechnology anti nqo 1 antibody
    L-malate (LM) increases expression of total Nrf2 and nuclear Nrf2 protein and HO-1, <t>NQO-1</t> protein after MIRI and decreases expression of Keap1 protein. Expression of total Nrf2, nuclear Nrf2, HO-1, NQO-1, and Keap1 was measured, with Western blot. Densitometric analysis was performed with Quantity One software 24 h later. Data are presented as means ± SD from 3 experiments. ∗∗ P
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    91
    Santa Cruz Biotechnology quinone 1
    Effects of safflower yellow B (SYB) on nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated antioxidant defense proteins in HepG2 cells. Cells were treated with SYB (50, 100 and 150 nmol/l) for 24 h prior to treatment with H 2 O 2 (200 µ mol/l). Total cell lysates were prepared and subjected to western blot analysis to monitor the expression levels of antioxidant proteins, namely Nrf2, heme oxygenase 1 (HO-1) and NAD(P)H dehydrogenase, <t>quinone</t> 1 (NQO1). β-actin served as an internal control. (A) Western blot analysis of Nrf2, HO-1 and NQO1 expression. Quantitative representations of western blot analysis of (B) Nrf2, (C) HO-1 and (D) NQ01. Data represent the means ± SD. * P
    Quinone 1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology rabbit anti nqo 1 antibody
    Effects of safflower yellow B (SYB) on nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated antioxidant defense proteins in HepG2 cells. Cells were treated with SYB (50, 100 and 150 nmol/l) for 24 h prior to treatment with H 2 O 2 (200 µ mol/l). Total cell lysates were prepared and subjected to western blot analysis to monitor the expression levels of antioxidant proteins, namely Nrf2, heme oxygenase 1 (HO-1) and NAD(P)H dehydrogenase, <t>quinone</t> 1 (NQO1). β-actin served as an internal control. (A) Western blot analysis of Nrf2, HO-1 and NQO1 expression. Quantitative representations of western blot analysis of (B) Nrf2, (C) HO-1 and (D) NQ01. Data represent the means ± SD. * P
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    Image Search Results


    BL153 upregulated renal PGC-1 α and NQO1 expression. PGC-1 α and NQO1 expression was detected by western blot assay. n = 5; * P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Magnolia Extract (BL153) Ameliorates Kidney Damage in a High Fat Diet-Induced Obesity Mouse Model

    doi: 10.1155/2013/367040

    Figure Lengend Snippet: BL153 upregulated renal PGC-1 α and NQO1 expression. PGC-1 α and NQO1 expression was detected by western blot assay. n = 5; * P

    Article Snippet: The latter were blocked with 5% milk, followed by incubation with the following antibodies: TNF-α , PGC-1α (Abcam, Cambridge, MA), PAI-1 (BD Bioscience, San Jose, CA), 3-NT (Millipore, Billerica, MA), 4-HNE (Alpha Diagnostic International, San Antonio, TX), HK II, β -actin, and NQO1 (SantaCruz Biotechnology, Santa Cruz, CA).

    Techniques: Pyrolysis Gas Chromatography, Expressing, Western Blot

    Effect on protein expression of Nrf2 targets. Relative Protein expression of GPX ( a ), HO-1 ( b ) and NQO1 ( c ) in the liver of rats fed diets with either soybean oil or salmon oil with various vitamin E concentrations. Bars represent relative protein level expressed as fold of control (diet with soybean oil and 25 mg/kg vitamin E) and are means ± SD from 12 animals per group. Representative immunoblots specific to GPX ( a ), HO-1 ( b ), NQO1 ( c ) and ß-actin as respective loading control are shown for one animal per group

    Journal: Nutrition & Metabolism

    Article Title: An excess dietary vitamin E concentration does not influence Nrf2 signaling in the liver of rats fed either soybean oil or salmon oil

    doi: 10.1186/s12986-017-0225-z

    Figure Lengend Snippet: Effect on protein expression of Nrf2 targets. Relative Protein expression of GPX ( a ), HO-1 ( b ) and NQO1 ( c ) in the liver of rats fed diets with either soybean oil or salmon oil with various vitamin E concentrations. Bars represent relative protein level expressed as fold of control (diet with soybean oil and 25 mg/kg vitamin E) and are means ± SD from 12 animals per group. Representative immunoblots specific to GPX ( a ), HO-1 ( b ), NQO1 ( c ) and ß-actin as respective loading control are shown for one animal per group

    Article Snippet: After that, the membranes were washed and blocked for 1 h at room temperature with 5% nonfat dry milk in TBS-T (w /v ) following incubations with primary antibodies against BIP (rabbit polyclonal anti-GRP78 antibody, Thermo Fisher Scientific Cat# PA5–29705, RRID:AB_2547179, Schwerte, Germany), p-eIF2α (rabbit polyclonal anti-pSer51 antibody, Cell Signaling Technology Cat# 9721, RRID:AB_330951, Frankfurt/Main, Germany), eIF2α (rabbit polyclonal anti-eIF2α antibody, Cell Signaling Technology Cat# 9722, RRID:AB_2230924), GPX (rabbit polyclonal anti-GPX antibody, Abcam Cat# ab22604, RRID:AB_2112120, Cambridge, UK), HO1 (rabbit polyclonal anti-HO1; Abcam Cat# ab68477, RRID:AB_11156457), Abcam), NQO1 (goat polyclonal, Santa Cruz Biotechnology Cat# sc-16,464, RRID:AB_2154339, Heidelberg, Germany) and β-actin (mouse monoclonal anti-β-actin, Abcam Cat# ab6276, RRID:AB_2223210, Abcam) or α-tubulin (rabbit monoclonal anti-α-tubulin, Cell Signaling Technology Cat# 2125, RRID:AB_2619646) as a reference protein for adequate normalization.

    Techniques: Expressing, Western Blot

    Hyperoxia increases pulmonary CYP1A1, NQO1, and MGST 1 protein expression

    Journal: Toxicology and applied pharmacology

    Article Title: Functional Deficiency of Aryl Hydrocarbon Receptor Augments Oxygen Toxicity-Induced Alveolar Simplification in Newborn Mice

    doi: 10.1016/j.taap.2013.01.003

    Figure Lengend Snippet: Hyperoxia increases pulmonary CYP1A1, NQO1, and MGST 1 protein expression

    Article Snippet: Thomas, Rutgers University, Piscataway, NJ, USA; dilution 1:1500), anti-NQO1 antibody (Santa Cruz Biotechnologies; sc-16464, dilution 1:500), anti-MGST1 antibody (Santa Cruz; sc-17003, dilution 1:500) and anti-β-actin antibody (Sigma-Aldrich; A5316, dilution 1:5000).

    Techniques: Expressing

    Hyperoxia increases pulmonary CYP1A1, NQO1, and MGST 1 mRNA expression

    Journal: Toxicology and applied pharmacology

    Article Title: Functional Deficiency of Aryl Hydrocarbon Receptor Augments Oxygen Toxicity-Induced Alveolar Simplification in Newborn Mice

    doi: 10.1016/j.taap.2013.01.003

    Figure Lengend Snippet: Hyperoxia increases pulmonary CYP1A1, NQO1, and MGST 1 mRNA expression

    Article Snippet: Thomas, Rutgers University, Piscataway, NJ, USA; dilution 1:1500), anti-NQO1 antibody (Santa Cruz Biotechnologies; sc-16464, dilution 1:500), anti-MGST1 antibody (Santa Cruz; sc-17003, dilution 1:500) and anti-β-actin antibody (Sigma-Aldrich; A5316, dilution 1:5000).

    Techniques: Expressing

    Hyperoxia increases pulmonary CYP1A1, NQO1, and GST enzyme activities

    Journal: Toxicology and applied pharmacology

    Article Title: Functional Deficiency of Aryl Hydrocarbon Receptor Augments Oxygen Toxicity-Induced Alveolar Simplification in Newborn Mice

    doi: 10.1016/j.taap.2013.01.003

    Figure Lengend Snippet: Hyperoxia increases pulmonary CYP1A1, NQO1, and GST enzyme activities

    Article Snippet: Thomas, Rutgers University, Piscataway, NJ, USA; dilution 1:1500), anti-NQO1 antibody (Santa Cruz Biotechnologies; sc-16464, dilution 1:500), anti-MGST1 antibody (Santa Cruz; sc-17003, dilution 1:500) and anti-β-actin antibody (Sigma-Aldrich; A5316, dilution 1:5000).

    Techniques:

    Kaplan-Meier survival curves illustrating the significance of NQO1 expression in ovarian carcinomas. (A) OS rates of patients with high (solid, n = 92) and low (dashed, n = 68) NQO1 expression. A: Log-rank = 21.699, P = 0.000. (B - C) High NQO1 expression was strongly associated with poor OS in early-stage (solid, n = 37) and late-stage (solid, n = 55). B: Log-rank = 6.527, P = 0.011; C: Log-rank = 4.806, P = 0.028. (D-F) High NQO1 expression was strongly associated with poor OS in G1 (solid, n = 10), G2 (solid, n = 29) and G3 (solid, n = 53). D: Log-rank = 4.359, P = 0.037; E: Log-rank = 7.020, P = 0.008; F: Log-rank = 5.978, P = 0.015).

    Journal: BMC Cancer

    Article Title: High expression of NQO1 is associated with poor prognosis in serous ovarian carcinoma

    doi: 10.1186/s12885-015-1271-4

    Figure Lengend Snippet: Kaplan-Meier survival curves illustrating the significance of NQO1 expression in ovarian carcinomas. (A) OS rates of patients with high (solid, n = 92) and low (dashed, n = 68) NQO1 expression. A: Log-rank = 21.699, P = 0.000. (B - C) High NQO1 expression was strongly associated with poor OS in early-stage (solid, n = 37) and late-stage (solid, n = 55). B: Log-rank = 6.527, P = 0.011; C: Log-rank = 4.806, P = 0.028. (D-F) High NQO1 expression was strongly associated with poor OS in G1 (solid, n = 10), G2 (solid, n = 29) and G3 (solid, n = 53). D: Log-rank = 4.359, P = 0.037; E: Log-rank = 7.020, P = 0.008; F: Log-rank = 5.978, P = 0.015).

    Article Snippet: The slides were then incubated with the NQO1 antibody (1:200, Santa Cruz Biotechnology, Dallas, TX, USA) at 4°C overnight.

    Techniques: Expressing

    IHC staining of NQO1 protein in ovarian tumor samples. (A) Negative expression of NQO1 protein in a benign serous tumor. (B–C) Weak positive expression of NQO1 protein (B) and positive expression (C) in atypical cells of borderline serous tumors. (D) Strong positive expression of NQO1 protein in serous carcinoma cells, in a patient with metastasis. (E) Positive expression of NQO1 protein in a serous carcinoma patient without metastasis. Scattered, strongly positive-staining cancer cells are seen ( arrows ). (F) Negative expression of NQO1 protein in a serous carcinoma patient without metastasis. Original magnification, A–F: ×200.

    Journal: BMC Cancer

    Article Title: High expression of NQO1 is associated with poor prognosis in serous ovarian carcinoma

    doi: 10.1186/s12885-015-1271-4

    Figure Lengend Snippet: IHC staining of NQO1 protein in ovarian tumor samples. (A) Negative expression of NQO1 protein in a benign serous tumor. (B–C) Weak positive expression of NQO1 protein (B) and positive expression (C) in atypical cells of borderline serous tumors. (D) Strong positive expression of NQO1 protein in serous carcinoma cells, in a patient with metastasis. (E) Positive expression of NQO1 protein in a serous carcinoma patient without metastasis. Scattered, strongly positive-staining cancer cells are seen ( arrows ). (F) Negative expression of NQO1 protein in a serous carcinoma patient without metastasis. Original magnification, A–F: ×200.

    Article Snippet: The slides were then incubated with the NQO1 antibody (1:200, Santa Cruz Biotechnology, Dallas, TX, USA) at 4°C overnight.

    Techniques: Immunohistochemistry, Staining, Expressing

    qRT-PCR analysis of NQO1 mRNA. Serous carcinoma specimens (n = 19) and benign serous tumors (n = 15) were collected, and NQO1 mRNA levels were assessed by qRT-PCR. Error bars represent the standard deviation of the mean (SD) calculated from three parallel experiments. ** P

    Journal: BMC Cancer

    Article Title: High expression of NQO1 is associated with poor prognosis in serous ovarian carcinoma

    doi: 10.1186/s12885-015-1271-4

    Figure Lengend Snippet: qRT-PCR analysis of NQO1 mRNA. Serous carcinoma specimens (n = 19) and benign serous tumors (n = 15) were collected, and NQO1 mRNA levels were assessed by qRT-PCR. Error bars represent the standard deviation of the mean (SD) calculated from three parallel experiments. ** P

    Article Snippet: The slides were then incubated with the NQO1 antibody (1:200, Santa Cruz Biotechnology, Dallas, TX, USA) at 4°C overnight.

    Techniques: Quantitative RT-PCR, Standard Deviation

    β -Lapachone induces programmed necrosis through NQO1 enzyme activity in SK-Hep1 cells. ( a ) SK-Hep1 cells stimulated with 4 μ M β -lapachone in the presence or absence of dicoumarol (50 μ M) for 18 h. Cell death was analyzed with LDH release. ( b ) SK-Hep1 cells were transfected with NQO1 siRNA or control siRNA. Twenty-four hours after transfection, cells were treated with the indicated concentrations of β -lapachone for 18 h. Western blotting analysis and LDH assay were performed. ( c ) Vector cells (SK-Hep1/Vector) and NQO1 overexpressed cells (SK-Hep1/NQO1) were treated with 1 μ M β -lapachone for 18 h. Cell death was analyzed using LDH assay (left panel). Western blotting analysis was performed for confirmed overexpression of NQO1 (right panel). ( d ) SK-Hep1 cells were transfected with vector, NQO1 and NQO1 shRNA. Twenty-four hours after transfection, cell were treated with 4 μ M β -lapachone for 1 h, and loaded with fluorescent dye, H 2 DCF-DA. H 2 DCF-DA fluorescence intensity was analyzed with a fluorescent microscope. The NQO1 protein levels were determined by western blotting. The level of actin was used as a loading control. ( e ) The NQO1 protein levels were determined by western blotting. The level of actin was used as a loading control (upper panel). NQO1 activity was determined in cell extracts prepared from U343, T98G, Caki, ACHN, MDA-MB-361, MDA-MB-231 and SK-Hep1 cells (lower panel). ( f , g ) U343, T98G, Caki, ACHN, MDA-MB-361, MDA-MB-231 and SK-Hep1 cells were treated with the indicated concentrations of β -lapachone for 14 h. Cell viability was analyzed using XTT assay ( f ). Cell death was analyzed with LDH assay ( g ). The values in a , b , c , e , f , g represent the mean±S.D. from three independent samples ( n =3). * P

    Journal: Cell Death & Disease

    Article Title: β-Lapachone induces programmed necrosis through the RIP1-PARP-AIF-dependent pathway in human hepatocellular carcinoma SK-Hep1 cells

    doi: 10.1038/cddis.2014.202

    Figure Lengend Snippet: β -Lapachone induces programmed necrosis through NQO1 enzyme activity in SK-Hep1 cells. ( a ) SK-Hep1 cells stimulated with 4 μ M β -lapachone in the presence or absence of dicoumarol (50 μ M) for 18 h. Cell death was analyzed with LDH release. ( b ) SK-Hep1 cells were transfected with NQO1 siRNA or control siRNA. Twenty-four hours after transfection, cells were treated with the indicated concentrations of β -lapachone for 18 h. Western blotting analysis and LDH assay were performed. ( c ) Vector cells (SK-Hep1/Vector) and NQO1 overexpressed cells (SK-Hep1/NQO1) were treated with 1 μ M β -lapachone for 18 h. Cell death was analyzed using LDH assay (left panel). Western blotting analysis was performed for confirmed overexpression of NQO1 (right panel). ( d ) SK-Hep1 cells were transfected with vector, NQO1 and NQO1 shRNA. Twenty-four hours after transfection, cell were treated with 4 μ M β -lapachone for 1 h, and loaded with fluorescent dye, H 2 DCF-DA. H 2 DCF-DA fluorescence intensity was analyzed with a fluorescent microscope. The NQO1 protein levels were determined by western blotting. The level of actin was used as a loading control. ( e ) The NQO1 protein levels were determined by western blotting. The level of actin was used as a loading control (upper panel). NQO1 activity was determined in cell extracts prepared from U343, T98G, Caki, ACHN, MDA-MB-361, MDA-MB-231 and SK-Hep1 cells (lower panel). ( f , g ) U343, T98G, Caki, ACHN, MDA-MB-361, MDA-MB-231 and SK-Hep1 cells were treated with the indicated concentrations of β -lapachone for 14 h. Cell viability was analyzed using XTT assay ( f ). Cell death was analyzed with LDH assay ( g ). The values in a , b , c , e , f , g represent the mean±S.D. from three independent samples ( n =3). * P

    Article Snippet: Anti-PARP-1, PAR, AIF and NQO1 antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Activity Assay, Transfection, Western Blot, Lactate Dehydrogenase Assay, Plasmid Preparation, Over Expression, shRNA, Fluorescence, Microscopy, Multiple Displacement Amplification, XTT Assay

    Reduced ROS generation and apoptosis resistance of BEAS2B-Cr-CSC. (A) Passage-matched normal BES-2B cells, BEAS-2B-Cr, and BEAS-2B-Cr-CSC were treated with various doses of Cr(VI) or cisplatin for 24 hours. Apoptosis was determined by Annexin V/PI using flow cytometry. (B) and (E) BEAS-2B, BEAS-2B-Cr, and BEAS-2B-Cr-CSC were harvested and whole protein lysates were isolated for examination of expressions of anti-apoptotic proteins Bcl-2 and Bcl-xL (B) and antioxidant enzymes SOD2 and NQO1 (E). The results are representative of three independent experiments. (C) Intracellular ROS levels were measured by DCFDA fluorescence using a fluorescence microplate reader. Data were normalized as compared with the ROS level in BEAS-2B cells without treatment. * and #, p

    Journal: Toxicology and applied pharmacology

    Article Title: Loss of fructose-1,6-bisphosphatase induces glycolysis and promotes apoptosis resistance of cancer stem-like cells: an important role in hexavalent chromium-induced carcinogenesis

    doi: 10.1016/j.taap.2017.06.014

    Figure Lengend Snippet: Reduced ROS generation and apoptosis resistance of BEAS2B-Cr-CSC. (A) Passage-matched normal BES-2B cells, BEAS-2B-Cr, and BEAS-2B-Cr-CSC were treated with various doses of Cr(VI) or cisplatin for 24 hours. Apoptosis was determined by Annexin V/PI using flow cytometry. (B) and (E) BEAS-2B, BEAS-2B-Cr, and BEAS-2B-Cr-CSC were harvested and whole protein lysates were isolated for examination of expressions of anti-apoptotic proteins Bcl-2 and Bcl-xL (B) and antioxidant enzymes SOD2 and NQO1 (E). The results are representative of three independent experiments. (C) Intracellular ROS levels were measured by DCFDA fluorescence using a fluorescence microplate reader. Data were normalized as compared with the ROS level in BEAS-2B cells without treatment. * and #, p

    Article Snippet: Antibodies against NQO1, GAPDH, and β-actin were obtained from Santa Cruz (Santa Cruz, CA).

    Techniques: Flow Cytometry, Cytometry, Isolation, Fluorescence

    mRNA expression of NQO1 . Comparison of NQO1 mRNA expression data assessed bycDNA microarray (y-axis) and quantitative reverse transcriptase PCR (x-axis). Microarray data are shown as log 10 -transformed, normalized ratios; qPCR data are shown as the log 10 -transformed, normalized starting quantities of the mRNA.

    Journal: BMC Cancer

    Article Title: ATBF1 and NQO1 as candidate targets for allelic loss at chromosome arm 16q in breast cancer: Absence of somatic ATBF1 mutations and no role for the C609T NQO1 polymorphism

    doi: 10.1186/1471-2407-8-105

    Figure Lengend Snippet: mRNA expression of NQO1 . Comparison of NQO1 mRNA expression data assessed bycDNA microarray (y-axis) and quantitative reverse transcriptase PCR (x-axis). Microarray data are shown as log 10 -transformed, normalized ratios; qPCR data are shown as the log 10 -transformed, normalized starting quantities of the mRNA.

    Article Snippet: The primary antibody recognized NQO1 (A180; Santa Cruz Biotechnology, Santa Cruz, CA), and it was used at a dilution of 1:1500.

    Techniques: Expressing, Microarray, Polymerase Chain Reaction, Transformation Assay, Real-time Polymerase Chain Reaction

    Immunohistochemical staining for NQO1 . A, normal, negative epithelium; B, negative tumor (0); C, weakly positive tumor (1); D, positive tumor (2).

    Journal: BMC Cancer

    Article Title: ATBF1 and NQO1 as candidate targets for allelic loss at chromosome arm 16q in breast cancer: Absence of somatic ATBF1 mutations and no role for the C609T NQO1 polymorphism

    doi: 10.1186/1471-2407-8-105

    Figure Lengend Snippet: Immunohistochemical staining for NQO1 . A, normal, negative epithelium; B, negative tumor (0); C, weakly positive tumor (1); D, positive tumor (2).

    Article Snippet: The primary antibody recognized NQO1 (A180; Santa Cruz Biotechnology, Santa Cruz, CA), and it was used at a dilution of 1:1500.

    Techniques: Immunohistochemistry, Staining

    NQO1 Activity of bone marrow cells of guinea pigs fed 0.5 mg or 15 mg vit C/day. (Panel A) AIR, exposed to air; DC, fed 3 mg DC/day; CS, exposed to CS; DC+CS, fed 3 mg DC/day and exposed to CS. * indicates significant difference (p

    Journal: PLoS ONE

    Article Title: NAD(P)H: Quinone Oxidoreductase 1 Deficiency Conjoint with Marginal Vitamin C Deficiency Causes Cigarette Smoke Induced Myelodysplastic Syndromes

    doi: 10.1371/journal.pone.0020590

    Figure Lengend Snippet: NQO1 Activity of bone marrow cells of guinea pigs fed 0.5 mg or 15 mg vit C/day. (Panel A) AIR, exposed to air; DC, fed 3 mg DC/day; CS, exposed to CS; DC+CS, fed 3 mg DC/day and exposed to CS. * indicates significant difference (p

    Article Snippet: NQO1 protein was measured by western blot followed by densitometry using antibody against NQO1 (Santa Cruz biotechnology, Inc., California, USA) as described elsewhere .

    Techniques: Activity Assay

    MDS produced in the guinea pigs are irreversible. (Panel A ) Differential staining showing persistent changes in blood and bone marrow cell morphology of MDS guinea pigs followed by discontinuation of CS exposure and feeding 15 mg vitamin C/day. A, a–d , represent sham controls (fed 0.5 mg vitamin C/day and exposed to air); A, e–h , represent MDS guinea pigs followed by discontinuation of CS exposure and feeding 15 mg vitamin C/day. Blood smear - Leishman stain; bone marrow aspirate - Wright Geimsa stain, except Perls' stain in d and h ; (magnification 400×). (Panel B ) Measurement of CD34(+) cells in bone marrow by flow cytometry. (Panel C ) Geimsa-stained metaphase spread showing aneuploidy in MDS guinea pigs followed by discontinuation of CS exposure and feeding 15 mg vitamin C/day; (magnification 1000×). (Panel D ) NQO1 Activity of bone marrow cells. Bars over the respective columns represent means ± SD (n = 4); * indicates significant difference (p

    Journal: PLoS ONE

    Article Title: NAD(P)H: Quinone Oxidoreductase 1 Deficiency Conjoint with Marginal Vitamin C Deficiency Causes Cigarette Smoke Induced Myelodysplastic Syndromes

    doi: 10.1371/journal.pone.0020590

    Figure Lengend Snippet: MDS produced in the guinea pigs are irreversible. (Panel A ) Differential staining showing persistent changes in blood and bone marrow cell morphology of MDS guinea pigs followed by discontinuation of CS exposure and feeding 15 mg vitamin C/day. A, a–d , represent sham controls (fed 0.5 mg vitamin C/day and exposed to air); A, e–h , represent MDS guinea pigs followed by discontinuation of CS exposure and feeding 15 mg vitamin C/day. Blood smear - Leishman stain; bone marrow aspirate - Wright Geimsa stain, except Perls' stain in d and h ; (magnification 400×). (Panel B ) Measurement of CD34(+) cells in bone marrow by flow cytometry. (Panel C ) Geimsa-stained metaphase spread showing aneuploidy in MDS guinea pigs followed by discontinuation of CS exposure and feeding 15 mg vitamin C/day; (magnification 1000×). (Panel D ) NQO1 Activity of bone marrow cells. Bars over the respective columns represent means ± SD (n = 4); * indicates significant difference (p

    Article Snippet: NQO1 protein was measured by western blot followed by densitometry using antibody against NQO1 (Santa Cruz biotechnology, Inc., California, USA) as described elsewhere .

    Techniques: Produced, Staining, Leishman Stain, Flow Cytometry, Cytometry, Activity Assay

    Exposure to permethrin (25 and 50 μ g/L) modules the protein levels of Nrf2-target antioxidant enzymes in zebrafish larvae. (a) Representative immunoblots, and densitometric analysis of immunoreactive bands of (b) γ -GCS (average for the control group = 0.52 γ -GSCs/ β -actin ratio), (c) HO-1 (average for the control group = 1.3 HO-1/ β -actin ratio), and (d) NQO-1 (average for the control group = 0.63 NQO-1/ β -actin ratio). The parametric dates are expressed as the mean ± SEM and analyzed by one-way ANOVA followed by Tukey as post hoc comparison, and nonparametric dates are expressed as median ± interquartile range, analyzed by Kruskal-Wallis followed by Dunn's multiple comparison test. Groups not sharing letters are significantly different ( p

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Acute Exposure to Permethrin Modulates Behavioral Functions, Redox, and Bioenergetics Parameters and Induces DNA Damage and Cell Death in Larval Zebrafish

    doi: 10.1155/2019/9149203

    Figure Lengend Snippet: Exposure to permethrin (25 and 50 μ g/L) modules the protein levels of Nrf2-target antioxidant enzymes in zebrafish larvae. (a) Representative immunoblots, and densitometric analysis of immunoreactive bands of (b) γ -GCS (average for the control group = 0.52 γ -GSCs/ β -actin ratio), (c) HO-1 (average for the control group = 1.3 HO-1/ β -actin ratio), and (d) NQO-1 (average for the control group = 0.63 NQO-1/ β -actin ratio). The parametric dates are expressed as the mean ± SEM and analyzed by one-way ANOVA followed by Tukey as post hoc comparison, and nonparametric dates are expressed as median ± interquartile range, analyzed by Kruskal-Wallis followed by Dunn's multiple comparison test. Groups not sharing letters are significantly different ( p

    Article Snippet: The membranes were blocked with 5% skimmed milk for 1 hour, washed in Tris-buffered saline with Tween-20 (100 mM Tris-HCl, pH 7.5, 0.9% NaCl, and 0.1% Tween-20), and incubated overnight at 4°C with the following rabbit primary antibodies: γ -GCS polyclonal antibody (rabbit anti-human; 1 : 1,000 dilution; cat. no. sc-22755; Santa Cruz Biotechnology), HO-1 (rabbit anti-human; 1 : 1,000 dilution; cat. no. sc-10789; Santa Cruz Biotechnology), NQO-1 (rabbit anti-human; 1 : 1,000 dilution; cat. no. sc-16464; Santa Cruz Biotechnology), and anti-β -actin polyclonal antibody (rabbit anti-human; 1 : 1,000 dilution; cat. no. A5060; Sigma-Aldrich).

    Techniques: Western Blot

    Reduced pyridine nucleotides and dicumarol induce a conformational change in NQO1. (A) Comparison of the ability of antibodies which bind to helix 7 (A180) and antibodies which bind to the C-terminal domain (C-Term) to immunoprecipitate rhNQO1 in the absence and presence of NADH. (B, C) Immunoprecipitation of rhNQO1 with the A180 antibody in the absence and presence of NADH or NADPH. (D) Immunoprecipitation of rhNQO1 by C-Term and A180 antibodies in the absence and presence of dicumarol. (E) The effect of NADH and dicumarol on the migration of rhNQO1 in non-denaturing PAGE. Reaction conditions for immunoprecipitation studies and non-denaturing PAGE are described in Materials and methods .

    Journal: PLoS ONE

    Article Title: Redox modulation of NQO1

    doi: 10.1371/journal.pone.0190717

    Figure Lengend Snippet: Reduced pyridine nucleotides and dicumarol induce a conformational change in NQO1. (A) Comparison of the ability of antibodies which bind to helix 7 (A180) and antibodies which bind to the C-terminal domain (C-Term) to immunoprecipitate rhNQO1 in the absence and presence of NADH. (B, C) Immunoprecipitation of rhNQO1 with the A180 antibody in the absence and presence of NADH or NADPH. (D) Immunoprecipitation of rhNQO1 by C-Term and A180 antibodies in the absence and presence of dicumarol. (E) The effect of NADH and dicumarol on the migration of rhNQO1 in non-denaturing PAGE. Reaction conditions for immunoprecipitation studies and non-denaturing PAGE are described in Materials and methods .

    Article Snippet: After 15 min either anti-NQO1 A180 or C-term antibody (2μg) was added for 1 h at 22°C followed by 25μl of protein A/G plus agarose (Santa Cruz Biotechnology) for an additional 30 min. Beads were collected by centrifugation at 5000 rpm for 1 min at 22°C, washed 3 times with 0.5ml of 25mM Tris-HCl, pH 7.4 containing 1mg/ml BSA and 5μM FAD, then once with 0.5ml of 25mM Tris-HCl, pH 7.4.

    Techniques: Immunoprecipitation, Migration, Polyacrylamide Gel Electrophoresis

    Increased immunostaining for NQO1 in 16HBE cells following treatment with β-lapachone. 16HBE cells were treated with DMSO or β-lapachone (10μM) for the indicated times then processed for immunocytochemistry and confocal analysis as described in Materials and Methods . Immunostaining for NQO1 was performed under non-denaturing conditions using the A180 antibody combined with DAPI nuclear staining.

    Journal: PLoS ONE

    Article Title: Redox modulation of NQO1

    doi: 10.1371/journal.pone.0190717

    Figure Lengend Snippet: Increased immunostaining for NQO1 in 16HBE cells following treatment with β-lapachone. 16HBE cells were treated with DMSO or β-lapachone (10μM) for the indicated times then processed for immunocytochemistry and confocal analysis as described in Materials and Methods . Immunostaining for NQO1 was performed under non-denaturing conditions using the A180 antibody combined with DAPI nuclear staining.

    Article Snippet: After 15 min either anti-NQO1 A180 or C-term antibody (2μg) was added for 1 h at 22°C followed by 25μl of protein A/G plus agarose (Santa Cruz Biotechnology) for an additional 30 min. Beads were collected by centrifugation at 5000 rpm for 1 min at 22°C, washed 3 times with 0.5ml of 25mM Tris-HCl, pH 7.4 containing 1mg/ml BSA and 5μM FAD, then once with 0.5ml of 25mM Tris-HCl, pH 7.4.

    Techniques: Immunostaining, Immunocytochemistry, Staining

    Relative positioning of the C-term and A180 antibody epitopes on human NQO1. Alternating viewpoints of the human NQO1 homodimer (PDB ID: 1D4A) with each monomer colored separately (blue and green) and the locations of the C-terminal epitopes (CT, yellow) and A180 epitopes (magenta) highlighted.

    Journal: PLoS ONE

    Article Title: Redox modulation of NQO1

    doi: 10.1371/journal.pone.0190717

    Figure Lengend Snippet: Relative positioning of the C-term and A180 antibody epitopes on human NQO1. Alternating viewpoints of the human NQO1 homodimer (PDB ID: 1D4A) with each monomer colored separately (blue and green) and the locations of the C-terminal epitopes (CT, yellow) and A180 epitopes (magenta) highlighted.

    Article Snippet: After 15 min either anti-NQO1 A180 or C-term antibody (2μg) was added for 1 h at 22°C followed by 25μl of protein A/G plus agarose (Santa Cruz Biotechnology) for an additional 30 min. Beads were collected by centrifugation at 5000 rpm for 1 min at 22°C, washed 3 times with 0.5ml of 25mM Tris-HCl, pH 7.4 containing 1mg/ml BSA and 5μM FAD, then once with 0.5ml of 25mM Tris-HCl, pH 7.4.

    Techniques:

    Curcumin inhibits NQO1 enzymatic activity in vitro and in cells. Enzymatic activity of recombinant NQO1 was determined in the absence (–) or presence (+) of BSA without (–) or with the indicated concentrations of curcumin ( A ) or dicoumarol

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Inhibition of NAD(P)H:quinone oxidoreductase 1 activity and induction of p53 degradation by the natural phenolic compound curcumin

    doi: 10.1073/pnas.0501828102

    Figure Lengend Snippet: Curcumin inhibits NQO1 enzymatic activity in vitro and in cells. Enzymatic activity of recombinant NQO1 was determined in the absence (–) or presence (+) of BSA without (–) or with the indicated concentrations of curcumin ( A ) or dicoumarol

    Article Snippet: The following Abs were used: monoclonal anti-p53 (Pab 240 and Pab 1801), goat anti-NQO1, rabbit anti-IκB (all from Santa Cruz Biotechnology), and monoclonal anti-Actin (Sigma).

    Techniques: Activity Assay, In Vitro, Recombinant

    Effects of ginsenoside Rd on HO-1 and NQO1 expression in rats subjected to myocardial I/R. The representative images of HO-1 and NQO1 are shown. The results are expressed as the mean ± S.E.M. (n = 8 in each group). #P

    Journal: International Journal of Clinical and Experimental Medicine

    Article Title: Ginsenoside Rd mitigates myocardial ischemia-reperfusion injury via Nrf2/HO-1 signaling pathway

    doi:

    Figure Lengend Snippet: Effects of ginsenoside Rd on HO-1 and NQO1 expression in rats subjected to myocardial I/R. The representative images of HO-1 and NQO1 are shown. The results are expressed as the mean ± S.E.M. (n = 8 in each group). #P

    Article Snippet: The membranes were then incubated with the primary antibody rabbit anti-Nrf2, rabbit anti-NQO1 (1:500, Santa Cruz Biotechnology) rabbit anti-HO-1 (1:2000, Santa Cruz Biotechnology).

    Techniques: Expressing

    Western blot analysis for the nuclear Nrf2, HO-1, and NQO1 protein levels 24 h after TBI and SFN treatment. (A−C) Representative photographs of the Western blot show Nr2, HO-1, and NQO1 protein levels in sham, vehicle and SFN-treatment animals,

    Journal: Acta Pharmacologica Sinica

    Article Title: The role of Nrf2 signaling in the regulation of antioxidants and detoxifying enzymes after traumatic brain injury in rats and mice

    doi: 10.1038/aps.2010.101

    Figure Lengend Snippet: Western blot analysis for the nuclear Nrf2, HO-1, and NQO1 protein levels 24 h after TBI and SFN treatment. (A−C) Representative photographs of the Western blot show Nr2, HO-1, and NQO1 protein levels in sham, vehicle and SFN-treatment animals,

    Article Snippet: Membranes were then incubated overnight with the primary antibody at the appropriate dilution (rabbit anti-Nrf2, 1:200, Santa Cruz Biotechnology; rabbit anti-HO-1, 1:2000, Epitomics, Burlingame, USA; rabbit anti-NQO1 antibody, 1:200, Santa Cruz Biotechnology).

    Techniques: Western Blot

    RT-PCR analysis for HO-1 and NQO1 mRNA levels at 24 h after TBI and SFN treatment. (A, B) Representative RT-PCR shows HO-1 and NQO1 mRNA levels in sham, vehicle and SFN-treated animals, respectively. (C, D) Quantification of HO-1 and NQO1 mRNA levels.

    Journal: Acta Pharmacologica Sinica

    Article Title: The role of Nrf2 signaling in the regulation of antioxidants and detoxifying enzymes after traumatic brain injury in rats and mice

    doi: 10.1038/aps.2010.101

    Figure Lengend Snippet: RT-PCR analysis for HO-1 and NQO1 mRNA levels at 24 h after TBI and SFN treatment. (A, B) Representative RT-PCR shows HO-1 and NQO1 mRNA levels in sham, vehicle and SFN-treated animals, respectively. (C, D) Quantification of HO-1 and NQO1 mRNA levels.

    Article Snippet: Membranes were then incubated overnight with the primary antibody at the appropriate dilution (rabbit anti-Nrf2, 1:200, Santa Cruz Biotechnology; rabbit anti-HO-1, 1:2000, Epitomics, Burlingame, USA; rabbit anti-NQO1 antibody, 1:200, Santa Cruz Biotechnology).

    Techniques: Reverse Transcription Polymerase Chain Reaction

    NQO1 inhibits PGC-1α proteasomal degradation and increases its protein half-life. (A) HEK-293T cells and HEK-293T cells stably expressing the Flag-tagged β4 (PSMB2) ring subunit were transiently transfected with untagged PGC-1α-

    Journal: Molecular and Cellular Biology

    Article Title: The Protein Level of PGC-1?, a Key Metabolic Regulator, Is Controlled by NADH-NQO1

    doi: 10.1128/MCB.01672-12

    Figure Lengend Snippet: NQO1 inhibits PGC-1α proteasomal degradation and increases its protein half-life. (A) HEK-293T cells and HEK-293T cells stably expressing the Flag-tagged β4 (PSMB2) ring subunit were transiently transfected with untagged PGC-1α-

    Article Snippet: The antibodies used were as follows: monoclonal mouse antiactin, anti-HA, anti-Flag (Sigma), mouse anti-NQO1 (Santa Cruz Biotechnology), mouse anti-PGC-1α (Calbiochem), polyclonal goat anti-NQO1 (Santa Cruz Biotechnology), polyclonal goat anti-NQO1 (ab2346; Abcam), and polyclonal rabbit anti-acetylated-Lys (9441; Cell Signaling).

    Techniques: Pyrolysis Gas Chromatography, Stable Transfection, Expressing, Transfection

    NQO1 knockdown in myoblasts reduces steady-state PGC-1α protein levels and activity. (A) NQO1 was knocked down in C2C12 cells using a lentivirus-based approach (3 individual experiments); all were run on the same gel, and cells were analyzed for

    Journal: Molecular and Cellular Biology

    Article Title: The Protein Level of PGC-1?, a Key Metabolic Regulator, Is Controlled by NADH-NQO1

    doi: 10.1128/MCB.01672-12

    Figure Lengend Snippet: NQO1 knockdown in myoblasts reduces steady-state PGC-1α protein levels and activity. (A) NQO1 was knocked down in C2C12 cells using a lentivirus-based approach (3 individual experiments); all were run on the same gel, and cells were analyzed for

    Article Snippet: The antibodies used were as follows: monoclonal mouse antiactin, anti-HA, anti-Flag (Sigma), mouse anti-NQO1 (Santa Cruz Biotechnology), mouse anti-PGC-1α (Calbiochem), polyclonal goat anti-NQO1 (Santa Cruz Biotechnology), polyclonal goat anti-NQO1 (ab2346; Abcam), and polyclonal rabbit anti-acetylated-Lys (9441; Cell Signaling).

    Techniques: Pyrolysis Gas Chromatography, Activity Assay

    NQO1 protects PGC-1α by NADH-dependent interaction. (A) HEK-293 cells were transfected as indicated and harvested for analysis 24 h posttransfection. HA beads were used to immunoprecipitate (IP) HA-tagged PGC-1α, whereas HA-p73β

    Journal: Molecular and Cellular Biology

    Article Title: The Protein Level of PGC-1?, a Key Metabolic Regulator, Is Controlled by NADH-NQO1

    doi: 10.1128/MCB.01672-12

    Figure Lengend Snippet: NQO1 protects PGC-1α by NADH-dependent interaction. (A) HEK-293 cells were transfected as indicated and harvested for analysis 24 h posttransfection. HA beads were used to immunoprecipitate (IP) HA-tagged PGC-1α, whereas HA-p73β

    Article Snippet: The antibodies used were as follows: monoclonal mouse antiactin, anti-HA, anti-Flag (Sigma), mouse anti-NQO1 (Santa Cruz Biotechnology), mouse anti-PGC-1α (Calbiochem), polyclonal goat anti-NQO1 (Santa Cruz Biotechnology), polyclonal goat anti-NQO1 (ab2346; Abcam), and polyclonal rabbit anti-acetylated-Lys (9441; Cell Signaling).

    Techniques: Pyrolysis Gas Chromatography, Transfection

    PGC-1α is an intrinsically disordered protein by prediction, susceptible to in vitro degradation by the 20S PC and protected by NQO1. (A) Analysis of PGC-1α and PCNA amino acid sequences by the FoldIndex prediction program. (B) In vitro

    Journal: Molecular and Cellular Biology

    Article Title: The Protein Level of PGC-1?, a Key Metabolic Regulator, Is Controlled by NADH-NQO1

    doi: 10.1128/MCB.01672-12

    Figure Lengend Snippet: PGC-1α is an intrinsically disordered protein by prediction, susceptible to in vitro degradation by the 20S PC and protected by NQO1. (A) Analysis of PGC-1α and PCNA amino acid sequences by the FoldIndex prediction program. (B) In vitro

    Article Snippet: The antibodies used were as follows: monoclonal mouse antiactin, anti-HA, anti-Flag (Sigma), mouse anti-NQO1 (Santa Cruz Biotechnology), mouse anti-PGC-1α (Calbiochem), polyclonal goat anti-NQO1 (Santa Cruz Biotechnology), polyclonal goat anti-NQO1 (ab2346; Abcam), and polyclonal rabbit anti-acetylated-Lys (9441; Cell Signaling).

    Techniques: Pyrolysis Gas Chromatography, In Vitro

    Schematic model illustrating the convergent actions of CREB, NQO1, AMPK, and Sirt1 on PGC-1α. The scheme summarizes PGC-1α transcription and posttranslational regulation by different known metabolite-sensing proteins, including NQO1.

    Journal: Molecular and Cellular Biology

    Article Title: The Protein Level of PGC-1?, a Key Metabolic Regulator, Is Controlled by NADH-NQO1

    doi: 10.1128/MCB.01672-12

    Figure Lengend Snippet: Schematic model illustrating the convergent actions of CREB, NQO1, AMPK, and Sirt1 on PGC-1α. The scheme summarizes PGC-1α transcription and posttranslational regulation by different known metabolite-sensing proteins, including NQO1.

    Article Snippet: The antibodies used were as follows: monoclonal mouse antiactin, anti-HA, anti-Flag (Sigma), mouse anti-NQO1 (Santa Cruz Biotechnology), mouse anti-PGC-1α (Calbiochem), polyclonal goat anti-NQO1 (Santa Cruz Biotechnology), polyclonal goat anti-NQO1 (ab2346; Abcam), and polyclonal rabbit anti-acetylated-Lys (9441; Cell Signaling).

    Techniques: Pyrolysis Gas Chromatography

    A role for NQO1 activity in regulating PGC-1α in fasting liver. (A) Mice were either fed ad libitum or fasted during the indicated time points (starting from the beginning of the night phase) and then sacrificed for hepatic analysis of NADH and

    Journal: Molecular and Cellular Biology

    Article Title: The Protein Level of PGC-1?, a Key Metabolic Regulator, Is Controlled by NADH-NQO1

    doi: 10.1128/MCB.01672-12

    Figure Lengend Snippet: A role for NQO1 activity in regulating PGC-1α in fasting liver. (A) Mice were either fed ad libitum or fasted during the indicated time points (starting from the beginning of the night phase) and then sacrificed for hepatic analysis of NADH and

    Article Snippet: The antibodies used were as follows: monoclonal mouse antiactin, anti-HA, anti-Flag (Sigma), mouse anti-NQO1 (Santa Cruz Biotechnology), mouse anti-PGC-1α (Calbiochem), polyclonal goat anti-NQO1 (Santa Cruz Biotechnology), polyclonal goat anti-NQO1 (ab2346; Abcam), and polyclonal rabbit anti-acetylated-Lys (9441; Cell Signaling).

    Techniques: Activity Assay, Pyrolysis Gas Chromatography, Mouse Assay

    NQO1 and PGC-1α steady-state levels in myoblasts and myotubes. (A) Analysis of PGC-1α and NQO1 protein and mRNA levels in C2C12 myoblast (MB) cells and at sequential days after the cells were put in differentiation medium. (B) Same as

    Journal: Molecular and Cellular Biology

    Article Title: The Protein Level of PGC-1?, a Key Metabolic Regulator, Is Controlled by NADH-NQO1

    doi: 10.1128/MCB.01672-12

    Figure Lengend Snippet: NQO1 and PGC-1α steady-state levels in myoblasts and myotubes. (A) Analysis of PGC-1α and NQO1 protein and mRNA levels in C2C12 myoblast (MB) cells and at sequential days after the cells were put in differentiation medium. (B) Same as

    Article Snippet: The antibodies used were as follows: monoclonal mouse antiactin, anti-HA, anti-Flag (Sigma), mouse anti-NQO1 (Santa Cruz Biotechnology), mouse anti-PGC-1α (Calbiochem), polyclonal goat anti-NQO1 (Santa Cruz Biotechnology), polyclonal goat anti-NQO1 (ab2346; Abcam), and polyclonal rabbit anti-acetylated-Lys (9441; Cell Signaling).

    Techniques: Pyrolysis Gas Chromatography

    NQO1 regulates PGC-1α accumulation in response to induction by starvation-mimicking conditions in mouse primary hepatocytes. (A) Primary hepatocytes were infected with either GFP- or NQO1-expressing adenoviruses. Expression levels of NQO1 mRNA

    Journal: Molecular and Cellular Biology

    Article Title: The Protein Level of PGC-1?, a Key Metabolic Regulator, Is Controlled by NADH-NQO1

    doi: 10.1128/MCB.01672-12

    Figure Lengend Snippet: NQO1 regulates PGC-1α accumulation in response to induction by starvation-mimicking conditions in mouse primary hepatocytes. (A) Primary hepatocytes were infected with either GFP- or NQO1-expressing adenoviruses. Expression levels of NQO1 mRNA

    Article Snippet: The antibodies used were as follows: monoclonal mouse antiactin, anti-HA, anti-Flag (Sigma), mouse anti-NQO1 (Santa Cruz Biotechnology), mouse anti-PGC-1α (Calbiochem), polyclonal goat anti-NQO1 (Santa Cruz Biotechnology), polyclonal goat anti-NQO1 (ab2346; Abcam), and polyclonal rabbit anti-acetylated-Lys (9441; Cell Signaling).

    Techniques: Pyrolysis Gas Chromatography, Infection, Expressing

    DPP3 overexpression promotes NRF2 nuclear accumulation and ROS resistance. ( A ) Levels of DPP3, NRF2 and NQO1 proteins in MCF7 cells stably overexpressing wt or mutant DPP3 proteins. GAPDH was used as a loading control. ( B ) Localization of overexpressed DPP3 and endogenous NRF2 in the MCF7 stable cell lines. Immunofluorescence was carried out using anti-HA and anti-NRF2 antibodies for DPP3 and NRF2, respectively. ( C ) ROS levels in the MCF7 stable cell lines. ( D-E ) Sensitivities of the MCF7 stable cell lines to H 2 O 2 ( D ) and diquat ( E ). Cells were treated with indicated concentrations of H 2 O 2 and diquat for 42 hr. Values presented are means from 2 independent experiments. Error bars represent standard deviations (SDs). Statistical significance was calculated by Student's t test comparing the values for the 2 ETGE mutants (T481E and G482E) with those of 3 ETGE-wt proteins (wt, Y318F and E451Q). *p

    Journal: Cancer research

    Article Title: NRF2 induction supporting breast cancer cell survival is enabled by oxidative stress-induced DPP3-KEAP1 interaction

    doi: 10.1158/0008-5472.CAN-16-2204

    Figure Lengend Snippet: DPP3 overexpression promotes NRF2 nuclear accumulation and ROS resistance. ( A ) Levels of DPP3, NRF2 and NQO1 proteins in MCF7 cells stably overexpressing wt or mutant DPP3 proteins. GAPDH was used as a loading control. ( B ) Localization of overexpressed DPP3 and endogenous NRF2 in the MCF7 stable cell lines. Immunofluorescence was carried out using anti-HA and anti-NRF2 antibodies for DPP3 and NRF2, respectively. ( C ) ROS levels in the MCF7 stable cell lines. ( D-E ) Sensitivities of the MCF7 stable cell lines to H 2 O 2 ( D ) and diquat ( E ). Cells were treated with indicated concentrations of H 2 O 2 and diquat for 42 hr. Values presented are means from 2 independent experiments. Error bars represent standard deviations (SDs). Statistical significance was calculated by Student's t test comparing the values for the 2 ETGE mutants (T481E and G482E) with those of 3 ETGE-wt proteins (wt, Y318F and E451Q). *p

    Article Snippet: The primary antibodies used are as follows: anti-DPP3 rabbit monoclonal (ab133671, Abcam), anti-NQO1 mouse monoclonal (sc-32793, Santa Cruz), anti-NRF2 rabbit monoclonal (ab62352, Abcam), anti-KEAP1 goat polyclonal (E20, sc-15246, Santa Cruz), anti-β-Actin mouse monoclonal (sc-69879, Santa Cruz), anti-GAPDH rabbit polyclonal (sc-25778, Santa Cruz) and anti-p62 rabbit monoclonal (ab109012, Abcam).

    Techniques: Over Expression, Stable Transfection, Mutagenesis, Immunofluorescence

    Effects of Nrf2 knockdown on cisplatin sensitivity in resistant A2780cp cells. A. After cells were transfected with Nrf2 siRNA (100 nM) for 48 h, cell lysates were collected for western blot analyses for Nrf2, Keap1, NQO1 and HO-1. B. Cells were transfected

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Nrf2 induces cisplatin resistance through activation of autophagy in ovarian carcinoma

    doi:

    Figure Lengend Snippet: Effects of Nrf2 knockdown on cisplatin sensitivity in resistant A2780cp cells. A. After cells were transfected with Nrf2 siRNA (100 nM) for 48 h, cell lysates were collected for western blot analyses for Nrf2, Keap1, NQO1 and HO-1. B. Cells were transfected

    Article Snippet: The membranes were incubated with rabbit monoclonal antibodies for Atg3, Atg5, beclin 1, Atg12 and Keap1 (1:1000) from Cell Signaling Technology; rabbit monoclonal antibody Nrf2 (1:20000) from Abcam; mouse monoclonal antibody NQO1 (1:2000) from Santa Cruz Biotechnology; rabbit polyclonal antibody p62 (1:500) from Proteintech; rabbit polyclonal antibody HO-1 (1:1000) from Enzo Life Science.

    Techniques: Transfection, Western Blot

    Protective effects of RSG against TS/EC-induced oxidative stress. a Immunofluorescence and Western blotting analysis emphasizing activation of the Nrf2 pathway. b Immunofluorescence and Western blotting analysis emphasizing the effect of RSG on activation of the transcription factor peroxisome proliferator-activated receptor (PPARγ). c Concurrently, the overexpression level of downstream detoxifying molecule NQO-1 in the cells including RSG was detected. n = 3 to 4 biological replicates. *p

    Journal: BMC Neuroscience

    Article Title: Assessing the protective effect of rosiglitazone against electronic cigarette/tobacco smoke-induced blood–brain barrier impairment

    doi: 10.1186/s12868-019-0497-5

    Figure Lengend Snippet: Protective effects of RSG against TS/EC-induced oxidative stress. a Immunofluorescence and Western blotting analysis emphasizing activation of the Nrf2 pathway. b Immunofluorescence and Western blotting analysis emphasizing the effect of RSG on activation of the transcription factor peroxisome proliferator-activated receptor (PPARγ). c Concurrently, the overexpression level of downstream detoxifying molecule NQO-1 in the cells including RSG was detected. n = 3 to 4 biological replicates. *p

    Article Snippet: The antibodies used in this study were purchased from the various sources: rabbit anti-PPARγ (#331500) from Invitrogen; rabbit anti-ZO-1 (#402200) from Life Technologies; rabbit anti-Nrf2 (#sc-722), mouse anti-NQO-1 (#sc-376023) and mouse anti-PECAM-1 (#sc-376764) from Santa Cruz Biotechnology.

    Techniques: Immunofluorescence, Western Blot, Activation Assay, Over Expression

    3g can elevate the protein level of Nrf2 and NQO1.

    Journal: Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry

    Article Title: Development of Novel Nrf2/ARE Inducers Bearing Pyrazino[2,1-a]isoquinolin Scaffold with Potent In Vitro Efficacy and Enhanced Physicochemical Properties

    doi: 10.3390/molecules22091541

    Figure Lengend Snippet: 3g can elevate the protein level of Nrf2 and NQO1.

    Article Snippet: Western Blot Anti-NQO1 (sc-271116), anti-Nrf2 (BS1258) and Anti-β-actin (AP0060) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and Bioworlde (Bioworlde, Visalia, CA, USA), respectively.

    Techniques:

    BR specifically inhibits the Nrf2 signaling pathway. A375 cells were treated with BR for 24 hours. (a) Cell survival was assessed using an MTS assay. (b and c) Cells were treated with various concentrations of BR for 24 hours ((b), right; (c) right) or with 50 nM BR for different time interval as indicated ((b), left; (c) left). Total cell extracts were prepared and subjected to Western blot using antibodies for Nrf2, HO-1, NQO1, GSTP1, I κ B α , COX2, NF- κ B, and actin. Data in (a) are shown as mean ± SD; ∗ P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: UVA Irradiation Enhances Brusatol-Mediated Inhibition of Melanoma Growth by Downregulation of the Nrf2-Mediated Antioxidant Response

    doi: 10.1155/2018/9742154

    Figure Lengend Snippet: BR specifically inhibits the Nrf2 signaling pathway. A375 cells were treated with BR for 24 hours. (a) Cell survival was assessed using an MTS assay. (b and c) Cells were treated with various concentrations of BR for 24 hours ((b), right; (c) right) or with 50 nM BR for different time interval as indicated ((b), left; (c) left). Total cell extracts were prepared and subjected to Western blot using antibodies for Nrf2, HO-1, NQO1, GSTP1, I κ B α , COX2, NF- κ B, and actin. Data in (a) are shown as mean ± SD; ∗ P

    Article Snippet: Nrf2 and NQO1 primary antibodies were purchased from Santa Cruz Biotechnology Inc. (USA).

    Techniques: MTS Assay, Western Blot

    Low-dose UVA can modulate the expression of phase II detoxification enzymes. (a) A375 cells were plated overnight and irradiated with different doses of UVA, and cell survival was determined 24 hours after UVA irradiation. (b and c) Cells were treated with UVA in the range of 0–100 kJ/m 2 for 24 hours. Total cell extracts were prepared for 24 hours following UVA irradiation and subjected to Western blot using antibodies for Nrf2, HO-1, NQO1, GSTP1, I κ B α , COX2, NF- κ B, and actin. We received a similar result in three independent experiments.

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: UVA Irradiation Enhances Brusatol-Mediated Inhibition of Melanoma Growth by Downregulation of the Nrf2-Mediated Antioxidant Response

    doi: 10.1155/2018/9742154

    Figure Lengend Snippet: Low-dose UVA can modulate the expression of phase II detoxification enzymes. (a) A375 cells were plated overnight and irradiated with different doses of UVA, and cell survival was determined 24 hours after UVA irradiation. (b and c) Cells were treated with UVA in the range of 0–100 kJ/m 2 for 24 hours. Total cell extracts were prepared for 24 hours following UVA irradiation and subjected to Western blot using antibodies for Nrf2, HO-1, NQO1, GSTP1, I κ B α , COX2, NF- κ B, and actin. We received a similar result in three independent experiments.

    Article Snippet: Nrf2 and NQO1 primary antibodies were purchased from Santa Cruz Biotechnology Inc. (USA).

    Techniques: Expressing, Irradiation, Western Blot

    Loss of NQO1 expression inhibits cell proliferation and in vivo tumor growth In A and B , A549 and H292 cells were assayed for proliferation rates at 0, 24, 48 and 72 h using the CyQuant cell proliferation kit (Life Technologies). In C , A549 shNQO1 cells (open symbols) and A549 shCtr-R (closed symbols) were subcutaneously injected into flanks of athymic mice at varying concentrations ((1.0 black), (2.5 blue) or (5.0 red) × 10 6 ) cells. Tumor growth was assessed bi/weekly using calipers. In D , A549 shNQO1 and A549 shCtr-R cells were injected subcutaneously into flanks of athymic mice and tumors were measured bi weekly by caliper measurements until a volume of 1000 mm 3 was reached. Kaplan-Meir survival analysis was conducted using GraphPad Prism 6 software. In E , representative photomicrograph of mice in C where mice were injected on the Left flank (L) with A549 shNQO1 cells or on the right flank (R) with A549 shCtr-R cells at the indication concentration of cells. Shown are mice whose tumors were photographed after 32 days. In F , Western Blot for tumor-NQO1 expression and PARP-1 cleavage. Samples were harvested in PARP-lysis buffer as described in “Materials and Methods”. In A , p values for A549 shNQO1 vs A549 shCtr-R cells at 24 h ( p = 0.0031), 48 h ( p

    Journal: Molecular cancer research : MCR

    Article Title: Depleting tumor-NQO1 Potentiates Anoikis and Inhibits Growth of NSCLC

    doi: 10.1158/1541-7786.MCR-15-0207-T

    Figure Lengend Snippet: Loss of NQO1 expression inhibits cell proliferation and in vivo tumor growth In A and B , A549 and H292 cells were assayed for proliferation rates at 0, 24, 48 and 72 h using the CyQuant cell proliferation kit (Life Technologies). In C , A549 shNQO1 cells (open symbols) and A549 shCtr-R (closed symbols) were subcutaneously injected into flanks of athymic mice at varying concentrations ((1.0 black), (2.5 blue) or (5.0 red) × 10 6 ) cells. Tumor growth was assessed bi/weekly using calipers. In D , A549 shNQO1 and A549 shCtr-R cells were injected subcutaneously into flanks of athymic mice and tumors were measured bi weekly by caliper measurements until a volume of 1000 mm 3 was reached. Kaplan-Meir survival analysis was conducted using GraphPad Prism 6 software. In E , representative photomicrograph of mice in C where mice were injected on the Left flank (L) with A549 shNQO1 cells or on the right flank (R) with A549 shCtr-R cells at the indication concentration of cells. Shown are mice whose tumors were photographed after 32 days. In F , Western Blot for tumor-NQO1 expression and PARP-1 cleavage. Samples were harvested in PARP-lysis buffer as described in “Materials and Methods”. In A , p values for A549 shNQO1 vs A549 shCtr-R cells at 24 h ( p = 0.0031), 48 h ( p

    Article Snippet: The process was repeated using a 1:5000 dilution of monoclonal NQO1 antibody (clone A-180, Santa Cruz Biotechnology).

    Techniques: Expressing, In Vivo, CyQUANT Assay, Injection, Mouse Assay, Software, Concentration Assay, Western Blot, Lysis

    Depleting NQO1 expression levels inhibits growth of NSCLC cells in soft agar In A , A549 shCtr-R and A549 shNQO1 cells were analyzed for their ability to form colonies and grow in soft agar. Photomicrographs shown are representative of experiments performed in sextuplet. In B- D , graphical representation of enumerated colonies for A549 shNQO1, A549 shNQO1-B and H292 shNQO1-B cells versus A549 shCtr-R, A549 shCtr-L and H292 shCtr-L cells. In B , p values for A549 shNQO1 vs A549 shCtr-R ( p (

    Journal: Molecular cancer research : MCR

    Article Title: Depleting tumor-NQO1 Potentiates Anoikis and Inhibits Growth of NSCLC

    doi: 10.1158/1541-7786.MCR-15-0207-T

    Figure Lengend Snippet: Depleting NQO1 expression levels inhibits growth of NSCLC cells in soft agar In A , A549 shCtr-R and A549 shNQO1 cells were analyzed for their ability to form colonies and grow in soft agar. Photomicrographs shown are representative of experiments performed in sextuplet. In B- D , graphical representation of enumerated colonies for A549 shNQO1, A549 shNQO1-B and H292 shNQO1-B cells versus A549 shCtr-R, A549 shCtr-L and H292 shCtr-L cells. In B , p values for A549 shNQO1 vs A549 shCtr-R ( p (

    Article Snippet: The process was repeated using a 1:5000 dilution of monoclonal NQO1 antibody (clone A-180, Santa Cruz Biotechnology).

    Techniques: Expressing

    Loss of NQO1 expression inhibits invasion of NSCLC In A , and C , 3D- tumor-spheroid invasion assays were performed on A549 shNQO1, H292 shNQO1-B and A549 and H292 shCtr-L cell lines as described in “Materials and Methods”. Shown are photomicrographs of representative spheroids from each cell line at 0 and 72 (h). In B and D , graphical presentation of the tumor-spheroid invasion distance migrated by A549 shNQO1 and H292 shNQO1-B cells as compared to A549 and H292 shCtr-L cell lines. In B , p values for A549 shNQO1 vs A549 shCtr-R (p

    Journal: Molecular cancer research : MCR

    Article Title: Depleting tumor-NQO1 Potentiates Anoikis and Inhibits Growth of NSCLC

    doi: 10.1158/1541-7786.MCR-15-0207-T

    Figure Lengend Snippet: Loss of NQO1 expression inhibits invasion of NSCLC In A , and C , 3D- tumor-spheroid invasion assays were performed on A549 shNQO1, H292 shNQO1-B and A549 and H292 shCtr-L cell lines as described in “Materials and Methods”. Shown are photomicrographs of representative spheroids from each cell line at 0 and 72 (h). In B and D , graphical presentation of the tumor-spheroid invasion distance migrated by A549 shNQO1 and H292 shNQO1-B cells as compared to A549 and H292 shCtr-L cell lines. In B , p values for A549 shNQO1 vs A549 shCtr-R (p

    Article Snippet: The process was repeated using a 1:5000 dilution of monoclonal NQO1 antibody (clone A-180, Santa Cruz Biotechnology).

    Techniques: Expressing

    Depleting tumor-NQO1 levels increases ROS formation and sensitizes NSCLC to anoikis In A and B , A549 shCtr-R, A549 shNQO1, H292 shCtr-L and H292 shNQO1-B cell lines were stained with 5 μM DCFDA ( Life technologies ) to detect endogenous ROS levels (H 2 O 2 ). In C and D cell death ELISA assays ( Roche Applied Sciences ) were performed on A549 and H292 NQO1 knockdown and control cells as described in “Methods” to detect cells that had undergone detachment induced cell death (anoikis). In A , p values for shCtr-R vs A549 shNQO1 ( p = 0.004). In B , p values for shCtr-L vs H292 shNQO1-B ( p = 0.0012). In C , p values for A549 shCtr-R vs A549 shNQO1 ( p

    Journal: Molecular cancer research : MCR

    Article Title: Depleting tumor-NQO1 Potentiates Anoikis and Inhibits Growth of NSCLC

    doi: 10.1158/1541-7786.MCR-15-0207-T

    Figure Lengend Snippet: Depleting tumor-NQO1 levels increases ROS formation and sensitizes NSCLC to anoikis In A and B , A549 shCtr-R, A549 shNQO1, H292 shCtr-L and H292 shNQO1-B cell lines were stained with 5 μM DCFDA ( Life technologies ) to detect endogenous ROS levels (H 2 O 2 ). In C and D cell death ELISA assays ( Roche Applied Sciences ) were performed on A549 and H292 NQO1 knockdown and control cells as described in “Methods” to detect cells that had undergone detachment induced cell death (anoikis). In A , p values for shCtr-R vs A549 shNQO1 ( p = 0.004). In B , p values for shCtr-L vs H292 shNQO1-B ( p = 0.0012). In C , p values for A549 shCtr-R vs A549 shNQO1 ( p

    Article Snippet: The process was repeated using a 1:5000 dilution of monoclonal NQO1 antibody (clone A-180, Santa Cruz Biotechnology).

    Techniques: Staining, Enzyme-linked Immunosorbent Assay

    NQO1 depletion causes a decrease in ALDH high activity In A , representative flow cytometry tracing of A549 shNQO1 and A549 shCtr-R cells analyzed for ALDH (high) activity using the Aldefluor Assay Kit from “Stem Cell Technologies”. Cells were assayed according to the manufacturers protocol as described in “Materials and Methods”. DEAB was used as an inhibitor of ALDH (high) activity. The percentages shown in each tracing indicate the population of cells staining for ALDH (high) activity. In B , graphical presentation of A549 shNQO1 and A549 shCtr-R cells assayed for ALDH (high) activity. The graph represents experiments repeated at least 5 times in duplicate.

    Journal: Molecular cancer research : MCR

    Article Title: Depleting tumor-NQO1 Potentiates Anoikis and Inhibits Growth of NSCLC

    doi: 10.1158/1541-7786.MCR-15-0207-T

    Figure Lengend Snippet: NQO1 depletion causes a decrease in ALDH high activity In A , representative flow cytometry tracing of A549 shNQO1 and A549 shCtr-R cells analyzed for ALDH (high) activity using the Aldefluor Assay Kit from “Stem Cell Technologies”. Cells were assayed according to the manufacturers protocol as described in “Materials and Methods”. DEAB was used as an inhibitor of ALDH (high) activity. The percentages shown in each tracing indicate the population of cells staining for ALDH (high) activity. In B , graphical presentation of A549 shNQO1 and A549 shCtr-R cells assayed for ALDH (high) activity. The graph represents experiments repeated at least 5 times in duplicate.

    Article Snippet: The process was repeated using a 1:5000 dilution of monoclonal NQO1 antibody (clone A-180, Santa Cruz Biotechnology).

    Techniques: Activity Assay, Flow Cytometry, Cytometry, Staining

    Elevated NQO1 levels in NSCLC patient tumors decreases their overall survival In A ). Patients were grouped into NQO1 low and NQO1 high expression groups as described in “Materials and Methods”. In B , Western blot analysis of A549 and H292 cells stably transduced with retroviral (shNQO1) or lentiviral (shNQO1-B) NQO1 constructs. In C and D , A549 and H292 NQO1 knockdown cell models from ( B ) were assayed for NQO1 enzyme activity, and activity was expressed as nMoles/min/μg of protein. In C , p values for A549 shCtr-R vs A549 shNQO1 ( p

    Journal: Molecular cancer research : MCR

    Article Title: Depleting tumor-NQO1 Potentiates Anoikis and Inhibits Growth of NSCLC

    doi: 10.1158/1541-7786.MCR-15-0207-T

    Figure Lengend Snippet: Elevated NQO1 levels in NSCLC patient tumors decreases their overall survival In A ). Patients were grouped into NQO1 low and NQO1 high expression groups as described in “Materials and Methods”. In B , Western blot analysis of A549 and H292 cells stably transduced with retroviral (shNQO1) or lentiviral (shNQO1-B) NQO1 constructs. In C and D , A549 and H292 NQO1 knockdown cell models from ( B ) were assayed for NQO1 enzyme activity, and activity was expressed as nMoles/min/μg of protein. In C , p values for A549 shCtr-R vs A549 shNQO1 ( p

    Article Snippet: The process was repeated using a 1:5000 dilution of monoclonal NQO1 antibody (clone A-180, Santa Cruz Biotechnology).

    Techniques: Expressing, Western Blot, Stable Transfection, Transduction, Construct, Activity Assay

    Expression of Nrf2, NQO1, MRP1, cmyc and p53 in CRC and their relationship to clinicopathologic variables

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Correlation of Nrf2, NQO1, MRP1, cmyc and p53 in colorectal cancer and their relationships to clinicopathologic features and survival

    doi:

    Figure Lengend Snippet: Expression of Nrf2, NQO1, MRP1, cmyc and p53 in CRC and their relationship to clinicopathologic variables

    Article Snippet: Monoclonal antibody against NQO1 (SC-271116) was purchased from Santa Cruz Biotechnology, Santa Cruz, CA.

    Techniques: Expressing

    Expression of Nrf2, NQO1, MRP1, cmyc and p53 was correlative and higher in CRC than adjacent non-tumor tissues

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Correlation of Nrf2, NQO1, MRP1, cmyc and p53 in colorectal cancer and their relationships to clinicopathologic features and survival

    doi:

    Figure Lengend Snippet: Expression of Nrf2, NQO1, MRP1, cmyc and p53 was correlative and higher in CRC than adjacent non-tumor tissues

    Article Snippet: Monoclonal antibody against NQO1 (SC-271116) was purchased from Santa Cruz Biotechnology, Santa Cruz, CA.

    Techniques: Expressing

    Association of Nrf2, NQO1, MRP1, cmyc and p53 expression with clinical prognosis

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Correlation of Nrf2, NQO1, MRP1, cmyc and p53 in colorectal cancer and their relationships to clinicopathologic features and survival

    doi:

    Figure Lengend Snippet: Association of Nrf2, NQO1, MRP1, cmyc and p53 expression with clinical prognosis

    Article Snippet: Monoclonal antibody against NQO1 (SC-271116) was purchased from Santa Cruz Biotechnology, Santa Cruz, CA.

    Techniques: Expressing

    Immunohistochemical staining results for Nrf2, NQO1, MRP1, cmyc and p53 expression in CRC. The unstained tissue slides from CRC were subject to HE staining (A1, A2) and IHC staining with antibodies against cmyc (B1, B2), MRP1 (C1, C2), NQO1 (D1, D2),

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Correlation of Nrf2, NQO1, MRP1, cmyc and p53 in colorectal cancer and their relationships to clinicopathologic features and survival

    doi:

    Figure Lengend Snippet: Immunohistochemical staining results for Nrf2, NQO1, MRP1, cmyc and p53 expression in CRC. The unstained tissue slides from CRC were subject to HE staining (A1, A2) and IHC staining with antibodies against cmyc (B1, B2), MRP1 (C1, C2), NQO1 (D1, D2),

    Article Snippet: Monoclonal antibody against NQO1 (SC-271116) was purchased from Santa Cruz Biotechnology, Santa Cruz, CA.

    Techniques: Immunohistochemistry, Staining, Expressing

    Expression of Nrf2, NQO1, MRP1, cmyc and p53 was correlative and higher in CRC than adjacent non-tumor tissues

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Correlation of Nrf2, NQO1, MRP1, cmyc and p53 in colorectal cancer and their relationships to clinicopathologic features and survival

    doi:

    Figure Lengend Snippet: Expression of Nrf2, NQO1, MRP1, cmyc and p53 was correlative and higher in CRC than adjacent non-tumor tissues

    Article Snippet: Monoclonal antibody against NQO1 (SC-271116) was purchased from Santa Cruz Biotechnology, Santa Cruz, CA.

    Techniques: Expressing

    Statistical analysis of IHC data using a quantitative scoring system. IHC staining sections were analyzed and measured on the Vectra platform to quantitate the expression of Nrf2, NQO1, MRP1, cmyc and p53 in both CRC tissues and adjacent non-tumor tissues.

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Correlation of Nrf2, NQO1, MRP1, cmyc and p53 in colorectal cancer and their relationships to clinicopathologic features and survival

    doi:

    Figure Lengend Snippet: Statistical analysis of IHC data using a quantitative scoring system. IHC staining sections were analyzed and measured on the Vectra platform to quantitate the expression of Nrf2, NQO1, MRP1, cmyc and p53 in both CRC tissues and adjacent non-tumor tissues.

    Article Snippet: Monoclonal antibody against NQO1 (SC-271116) was purchased from Santa Cruz Biotechnology, Santa Cruz, CA.

    Techniques: Immunohistochemistry, Staining, Expressing

    L-malate (LM) increases expression of total Nrf2 and nuclear Nrf2 protein and HO-1, NQO-1 protein after MIRI and decreases expression of Keap1 protein. Expression of total Nrf2, nuclear Nrf2, HO-1, NQO-1, and Keap1 was measured, with Western blot. Densitometric analysis was performed with Quantity One software 24 h later. Data are presented as means ± SD from 3 experiments. ∗∗ P

    Journal: Evidence-based Complementary and Alternative Medicine : eCAM

    Article Title: Protective Effects of L-Malate against Myocardial Ischemia/Reperfusion Injury in Rats

    doi: 10.1155/2016/3803657

    Figure Lengend Snippet: L-malate (LM) increases expression of total Nrf2 and nuclear Nrf2 protein and HO-1, NQO-1 protein after MIRI and decreases expression of Keap1 protein. Expression of total Nrf2, nuclear Nrf2, HO-1, NQO-1, and Keap1 was measured, with Western blot. Densitometric analysis was performed with Quantity One software 24 h later. Data are presented as means ± SD from 3 experiments. ∗∗ P

    Article Snippet: Antibodies for actin (number sc-1616r), anti-Nrf2 (number sc-722), anti-HO-1 (sc-1797), and anti-NQO-1 (sc-25591) were from Santa Cruz Biotechnology Inc. (Santa Cruz, CA), and anti-Keap1 (number bs6783) was from Bio world Technology Inc. (St Louis Park, MN).

    Techniques: Expressing, Western Blot, Software

    Effects of safflower yellow B (SYB) on nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated antioxidant defense proteins in HepG2 cells. Cells were treated with SYB (50, 100 and 150 nmol/l) for 24 h prior to treatment with H 2 O 2 (200 µ mol/l). Total cell lysates were prepared and subjected to western blot analysis to monitor the expression levels of antioxidant proteins, namely Nrf2, heme oxygenase 1 (HO-1) and NAD(P)H dehydrogenase, quinone 1 (NQO1). β-actin served as an internal control. (A) Western blot analysis of Nrf2, HO-1 and NQO1 expression. Quantitative representations of western blot analysis of (B) Nrf2, (C) HO-1 and (D) NQ01. Data represent the means ± SD. * P

    Journal: International Journal of Molecular Medicine

    Article Title: Safflower yellow B suppresses HepG2 cell injury induced by oxidative stress through the AKT/Nrf2 pathway

    doi: 10.3892/ijmm.2016.2462

    Figure Lengend Snippet: Effects of safflower yellow B (SYB) on nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated antioxidant defense proteins in HepG2 cells. Cells were treated with SYB (50, 100 and 150 nmol/l) for 24 h prior to treatment with H 2 O 2 (200 µ mol/l). Total cell lysates were prepared and subjected to western blot analysis to monitor the expression levels of antioxidant proteins, namely Nrf2, heme oxygenase 1 (HO-1) and NAD(P)H dehydrogenase, quinone 1 (NQO1). β-actin served as an internal control. (A) Western blot analysis of Nrf2, HO-1 and NQO1 expression. Quantitative representations of western blot analysis of (B) Nrf2, (C) HO-1 and (D) NQ01. Data represent the means ± SD. * P

    Article Snippet: Anti-β-actin primary (sc-7210), anti-AKT (sc-8312), anti-phosphorylated (p-)AKT (sc-33437), anti-heme oxygenase 1 (HO-1; sc-10789), anti-nuclear factor erythroid 2-related factor 2 (Nrf2; sc-7943) and anti-NAD(P)H dehydrogenase, quinone 1 (NQO1; sc-25591) antibodies were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

    Techniques: Western Blot, Expressing