anti-nrf2 Search Results


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  • 99
    Abcam rb anti nrf 2
    Rb Anti Nrf 2, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam antinrf2
    Antinrf2, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam anti nrf 2
    Effects of DJ-1 on expression of <t>Nrf2.</t> a After OGD for 5 h followed by 24 h of reoxygenation, astrocytes were harvested. Western blot for Nrf2 and CRIF1 in astrocytes. b After 24 h of reperfusion, brains were collected. Western blot for Nrf2 and CRIF1 in rats. c , d Ratios of Nrf2 and CRIF1 relative to β-actin in astrocytes and in rats, respectively. e Immunocytochemistry assays to measure total Nrf2 and nuclear Nrf2 in astrocytes. Fluorescence microscopy was used to assess expression. Nrf2 expression in astrocytes is indicated by red fluorescence. Cell nuclei were stained with DAPI. Original magnification, 200×. f Semi-quantitation to determine total number of Nrf2-positive cells and number of nuclear Nrf2-positive cells in astrocytes. Values are mean ± SEM. # p
    Anti Nrf 2, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 99 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti nrf 2/product/Abcam
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    93
    Santa Cruz Biotechnology anti nrf2
    Protective effect of eriodictyol-induced HO-1 expression on H 2 O 2 -induced cell damage. ( A ) H 2 O 2 -exposed cells were pretreated for 1 h with or without ZnPP and then treated with eriodictyol. The inhibitory effect of eriodictyol on H 2 O 2 -induced intracellular ROS generation was observed by fluorescence microscopy; ( B ) H 2 O 2 -stimulated cells were pretreated for 1 h with or without ZnPP or <t>Nrf2</t> or HO-1 siRNA and then treated with eriodictyol. Protective effect of HO-1 induction on cell death was determined by in situ terminal nick end-labeling (TUNEL) assay: −, untreated; +, treated. ROS and TUNEL staining was quantified in four randomly selected fields for each group: C, control. The scale bars for image ( B ) are the same as that in ( A ), 100 μm. * p
    Anti Nrf2, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1542 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    GeneTex anti nrf2
    Andrographolide induces <t>Nrf2</t> expression in Ava5 cells. (A–C) Induction of Nrf2 expression (A), Nrf2 nuclear translocation (B) and Nrf2-mediated ARE transactivation (C) by andrographolide in a concentration- or time-dependent manner. Ava5 or p2xARE-Luc-transfected
    Anti Nrf2, supplied by GeneTex, used in various techniques. Bioz Stars score: 92/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Epitomics anti nrf2
    DJ-1 does not impact the interaction between <t>Nrf2</t> and Keap1. ( A ) DJ-1 does not interact with Nrf2. HeLa cells were transfected with Flag-Nrf2 and Myc-DJ-1 for 48 h. Cells were washed twice with PBS, and dithiobis[succinimidylpropionate] (DSP, Thermo Scientific)
    Anti Nrf2, supplied by Epitomics, used in various techniques. Bioz Stars score: 92/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti nrf2/product/Epitomics
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    97
    Cell Signaling Technology Inc nrf2
    Probucol activated the <t>Nrf2/ARE</t> signaling pathways in astrocytes a . b . Immunofluorescence analysis was used to detect the staining intensity of GFAP and Nrf2. Scale bars are 50 μm. * P
    Nrf2, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 1118 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Millipore nrf2
    Immunoblotting of <t>NRF2,</t> AMPK, HIF, evaluated on HS-68 cells exposed to 1 μmol/L BDE 209, 99, 47 and MIX for 12 and 20 days. Actin was used as internal control. The images are representative of at least three separate experiments. The relative protein quantification is represented in the graphic (* p
    Nrf2, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 252 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Proteintech nrf2
    The protective effects of the H 2 S donor on the expression of antioxidant-related proteins in the ageing kidney tissue. (a) The expression change of <t>Nrf2</t> ( N = 11), HO-1 ( N = 10), SOD1 ( N = 11), and SOD2 ( N = 10) after 10 weeks of exogenous H 2 S donor. (b) Nrf2 was translocated from cytosol to nucleus, Lamin B was used as nuclear control, and GAPDH was used as cytosol control ( N = 6). Values are mean ± SE. P
    Nrf2, supplied by Proteintech, used in various techniques. Bioz Stars score: 95/100, based on 136 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher nrf2
    Klotho improved nuclear translocation of <t>Nrf2</t> in DOX-exposed myocardium and myocytes which was reversed by MAPKs inhibitors or siRNAs. ( A, B ) The left panels show the immunoblots of Nrf2 and Histone H3 in nuclear protein samples extracted from myocardium and myocytes. Columns on the right panels indicate the ratio of Nrf2/histone H3 in myocardium and cultured myocytes respectively. ( C ) The left panel demonstrates the captured fluorescent images of Nrf2, DAPI, and their merged images in cultured myocytes. Columns on the right part indicate the Nrf2 nuclear translocation rate in different groups. a Differences were significant when compared with control ( p
    Nrf2, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 282 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    ABclonal anti nrf2
    Schematic representation of the p62- Keap1- <t>Nrf2</t> pathway in ovarian cancer cells with the treatment of VK3. Our study provides evidence that p62 promotes Nrf2 signaling through interacting with Keap1, which blocks VK3-induced apoptosis by inhibiting ROS production in SKOV3/DDP cells.
    Anti Nrf2, supplied by ABclonal, used in various techniques. Bioz Stars score: 92/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    nrf2  (Bioss)
    92
    Bioss nrf2
    Effects of dietary resveratrol supplementation during gestation and lactation of sows on mRNA relative expression of <t>Nrf2-regulated</t> genes and pro-inflammatory cytokines in placenta. a , b mRNA relative expression of Nrf2-regulated genes; c mRNA relative expression of pro-inflammatory cytokines. Con, control treatment; Res, resveratrol treatment; SOD , superoxide dismutase; Gpx , glutathione peroxidase; CAT , catalase; NQO1 , NAD(P)H quinone dehydrogenase 1; HO1 , heme oxygenase 1; CYP1A1 , cytochrome P450 family 1 subfamily A member 1; TXNRD1 , thioredoxin reductase 1; GCLM , glutamate-cysteine ligase modifier; MGST1 , microsomal glutathione S-transferase 1; UGT1A1 , UDP glucuronosyl-transferase family 1 member A1; IL-1β , interleukin 1β; IL-6 , interleukin 6; IL-8 , interleukin 8; TNF-α , tumor necrosis factor α. All values are expressed as means ± SEM (n = 6). GAPDH was used as the internal control. * P
    Nrf2, supplied by Bioss, used in various techniques. Bioz Stars score: 92/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Enzo Biochem nrf2
    DHA enhances the nuclear translocation and DNA binding activity of <t>Nrf2</t> in neurons. Primary neuronal cultures were treated with vehicle or DHA followed by OGD. A , B , Representative Western blots ( A ) and semiquantitative analyses ( B ) of Nrf2 levels, showing increased nuclear Nrf2 in DHA-treated groups (* p ≤ 0.05 vs control and vehicle-treated OGD groups at the same time points; # p ≤ 0.05 vs control groups, n = 3 per condition). C , D , Representative electrophoretic mobility shift assay ( C ) and semiquantitative analyses ( D ) of DNA binding of Nrf2 (* p ≤ 0.05 vs control and vehicle-treated OGD groups at the same time points; # p ≤ 0.05 vs control groups; and p ≤ 0.01 vs DHA-treated OGD at 24 h; n = 3 per condition). The specificity of DNA binding activity was determined by a competition assay, whereas threefold (C1) or 50-fold (C2) excess of cold probe was added as a specific competitor.
    Nrf2, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 92/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    OriGene anti nrf2
    Ethanol induces <t>Nrf2/ARE</t> binding activity in PCN nuclear extracts. A, nuclear protein obtained from controls and PCNs exposed to ETOH (4 mg/ml) for different time points was used to detect ARE-specific oligonucleotide-protein complexes by EMSA. B, supershift
    Anti Nrf2, supplied by OriGene, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Effects of DJ-1 on expression of Nrf2. a After OGD for 5 h followed by 24 h of reoxygenation, astrocytes were harvested. Western blot for Nrf2 and CRIF1 in astrocytes. b After 24 h of reperfusion, brains were collected. Western blot for Nrf2 and CRIF1 in rats. c , d Ratios of Nrf2 and CRIF1 relative to β-actin in astrocytes and in rats, respectively. e Immunocytochemistry assays to measure total Nrf2 and nuclear Nrf2 in astrocytes. Fluorescence microscopy was used to assess expression. Nrf2 expression in astrocytes is indicated by red fluorescence. Cell nuclei were stained with DAPI. Original magnification, 200×. f Semi-quantitation to determine total number of Nrf2-positive cells and number of nuclear Nrf2-positive cells in astrocytes. Values are mean ± SEM. # p

    Journal: Journal of Molecular Medicine (Berlin, Germany)

    Article Title: Effect of DJ-1 on the neuroprotection of astrocytes subjected to cerebral ischemia/reperfusion injury

    doi: 10.1007/s00109-018-1719-5

    Figure Lengend Snippet: Effects of DJ-1 on expression of Nrf2. a After OGD for 5 h followed by 24 h of reoxygenation, astrocytes were harvested. Western blot for Nrf2 and CRIF1 in astrocytes. b After 24 h of reperfusion, brains were collected. Western blot for Nrf2 and CRIF1 in rats. c , d Ratios of Nrf2 and CRIF1 relative to β-actin in astrocytes and in rats, respectively. e Immunocytochemistry assays to measure total Nrf2 and nuclear Nrf2 in astrocytes. Fluorescence microscopy was used to assess expression. Nrf2 expression in astrocytes is indicated by red fluorescence. Cell nuclei were stained with DAPI. Original magnification, 200×. f Semi-quantitation to determine total number of Nrf2-positive cells and number of nuclear Nrf2-positive cells in astrocytes. Values are mean ± SEM. # p

    Article Snippet: Immunocytochemistry and immunohistochemistry Cells grown on glass coverslips and frozen sections of brain tissue were lightly fixed in 4% paraformaldehyde, blocked with 3% FBS/0.01% Triton X-100 in PBS, and incubated at 4 °C overnight with the following primary antibodies: anti-DJ-1 (1:50, Abcam, ab76008), anti-Nrf2 (1:50, Abcam, ab31163), and anti-GFAP (1:200, ABclonal, A10871).

    Techniques: Expressing, Western Blot, Immunocytochemistry, Fluorescence, Microscopy, Staining, Quantitation Assay

    DJ-1 regulates Nrf2/ARE binding activity and Nrf2/ARE-driven gene expression. After OGD for 5 h followed by 24 h of reoxygenation, astrocytes were harvested. And after 24 h of reperfusion, brains were collected. a EMSA analysis of Nrf2/ARE binding. b Semiquantitative analysis of Nrf2/ARE binding. CK, 100x, (+) and (−) indicate controls. c Western blot for GCLM, GCLC, and GSS in astrocytes. d Western blot for GCLM, GCLC, and GSS in rats. e , f Ratios of GCLM, GCLC, and GSS relative to β-actin in astrocytes and in rats, respectively. Values are mean ± SEM. # p

    Journal: Journal of Molecular Medicine (Berlin, Germany)

    Article Title: Effect of DJ-1 on the neuroprotection of astrocytes subjected to cerebral ischemia/reperfusion injury

    doi: 10.1007/s00109-018-1719-5

    Figure Lengend Snippet: DJ-1 regulates Nrf2/ARE binding activity and Nrf2/ARE-driven gene expression. After OGD for 5 h followed by 24 h of reoxygenation, astrocytes were harvested. And after 24 h of reperfusion, brains were collected. a EMSA analysis of Nrf2/ARE binding. b Semiquantitative analysis of Nrf2/ARE binding. CK, 100x, (+) and (−) indicate controls. c Western blot for GCLM, GCLC, and GSS in astrocytes. d Western blot for GCLM, GCLC, and GSS in rats. e , f Ratios of GCLM, GCLC, and GSS relative to β-actin in astrocytes and in rats, respectively. Values are mean ± SEM. # p

    Article Snippet: Immunocytochemistry and immunohistochemistry Cells grown on glass coverslips and frozen sections of brain tissue were lightly fixed in 4% paraformaldehyde, blocked with 3% FBS/0.01% Triton X-100 in PBS, and incubated at 4 °C overnight with the following primary antibodies: anti-DJ-1 (1:50, Abcam, ab76008), anti-Nrf2 (1:50, Abcam, ab31163), and anti-GFAP (1:200, ABclonal, A10871).

    Techniques: Binding Assay, Activity Assay, Expressing, Western Blot

    Mechanisms of action of DJ-1 in astrocytes. After oxidative stress induced by cerebral ischemia and reperfusion, DJ-1 is expressed in immunoreactive astrocytes. DJ-1 facilitates Nrf2 translocation to the nucleus by preventing binding with Keap1. Nrf2 binds to ARE and upregulates expression of GCLC, GCLM, and GSS, which regulate synthesis of GSH

    Journal: Journal of Molecular Medicine (Berlin, Germany)

    Article Title: Effect of DJ-1 on the neuroprotection of astrocytes subjected to cerebral ischemia/reperfusion injury

    doi: 10.1007/s00109-018-1719-5

    Figure Lengend Snippet: Mechanisms of action of DJ-1 in astrocytes. After oxidative stress induced by cerebral ischemia and reperfusion, DJ-1 is expressed in immunoreactive astrocytes. DJ-1 facilitates Nrf2 translocation to the nucleus by preventing binding with Keap1. Nrf2 binds to ARE and upregulates expression of GCLC, GCLM, and GSS, which regulate synthesis of GSH

    Article Snippet: Immunocytochemistry and immunohistochemistry Cells grown on glass coverslips and frozen sections of brain tissue were lightly fixed in 4% paraformaldehyde, blocked with 3% FBS/0.01% Triton X-100 in PBS, and incubated at 4 °C overnight with the following primary antibodies: anti-DJ-1 (1:50, Abcam, ab76008), anti-Nrf2 (1:50, Abcam, ab31163), and anti-GFAP (1:200, ABclonal, A10871).

    Techniques: Translocation Assay, Binding Assay, Expressing

    Schematic model for the induction of aggresome formation by NRF2 during proteasome inhibition. Under physiological conditions, Nrf2 is degraded by the Ub proteasome system. In cells with reduced proteasome activity, Nrf2 is stabilized and accumulates in the nuclei. Through transcriptional activation of p62, Nrf2 promotes aggresomes formation. Through functioning as a sensor of cytosolic proteasome activity and an activator of aggresome formation, Nrf2 alleviates cell damages caused by proteasomal stress. Nrf2, nuclear factor erythroid 2-related factor 2; ARE, antioxidant response elements; Ub, ubiquitin.

    Journal: Biomedical Reports

    Article Title: Transcriptional factor Nrf2 is essential for aggresome formation during proteasome inhibition

    doi: 10.3892/br.2019.1247

    Figure Lengend Snippet: Schematic model for the induction of aggresome formation by NRF2 during proteasome inhibition. Under physiological conditions, Nrf2 is degraded by the Ub proteasome system. In cells with reduced proteasome activity, Nrf2 is stabilized and accumulates in the nuclei. Through transcriptional activation of p62, Nrf2 promotes aggresomes formation. Through functioning as a sensor of cytosolic proteasome activity and an activator of aggresome formation, Nrf2 alleviates cell damages caused by proteasomal stress. Nrf2, nuclear factor erythroid 2-related factor 2; ARE, antioxidant response elements; Ub, ubiquitin.

    Article Snippet: The following primary antibodies were used: Anti-Nrf2 (cat. no. ab137550; 1:2,000; Abcam), anti-p62 (cat.no.

    Techniques: Inhibition, Activity Assay, Activation Assay

    Expression of p62 rescues the defects of proteasomal stress response in Nrf2 knockout cells. (A) Representative images and quantification of aggresomes in nrf2 +/+ , nrf2 -/- [Vector], nrf2 -/- [Flag-Nrf2] and nrf2 -/- [Flag-p62] cells after 14 h treatment with MG132 (2 µM). Anti-UbK48 was used to visualize ubiquitin-containing aggresomes (arrowheads), anti-Flag was used to visualize exogenous Nrf2 and p62, and DAPI was used to detect nuclei; scale bar, 20 µm. (B) Representative images and quantification of nrf2 +/+ , nrf2 -/- [Vector], nrf2 -/- [Flag-Nrf2] and nrf2 -/- [Flag-p62] cells stained with Annexin V (green) and PI (red) after 20 h treatment with DMSO or MG132. Live cells, Annexin V - PI - ; early apoptotic cells, Annexin V + PI - ; necrotic cells, Annexin V - PI + ; late apoptotic cells, Annexin V + PI + ; scale bar, 100 µm. (C) Representative immunoblots and quantification of p62 and Nrf2 levels in nrf2 +/+ , nrf2 -/- [Vector], nrf2 -/- [Flag-Nrf2] and nrf2 -/- [Flag-p62]. Data are presented as the mean ± SEM representative of three independent experiments and were assessed using one-way ANOVA followed by Tukey's test. * P

    Journal: Biomedical Reports

    Article Title: Transcriptional factor Nrf2 is essential for aggresome formation during proteasome inhibition

    doi: 10.3892/br.2019.1247

    Figure Lengend Snippet: Expression of p62 rescues the defects of proteasomal stress response in Nrf2 knockout cells. (A) Representative images and quantification of aggresomes in nrf2 +/+ , nrf2 -/- [Vector], nrf2 -/- [Flag-Nrf2] and nrf2 -/- [Flag-p62] cells after 14 h treatment with MG132 (2 µM). Anti-UbK48 was used to visualize ubiquitin-containing aggresomes (arrowheads), anti-Flag was used to visualize exogenous Nrf2 and p62, and DAPI was used to detect nuclei; scale bar, 20 µm. (B) Representative images and quantification of nrf2 +/+ , nrf2 -/- [Vector], nrf2 -/- [Flag-Nrf2] and nrf2 -/- [Flag-p62] cells stained with Annexin V (green) and PI (red) after 20 h treatment with DMSO or MG132. Live cells, Annexin V - PI - ; early apoptotic cells, Annexin V + PI - ; necrotic cells, Annexin V - PI + ; late apoptotic cells, Annexin V + PI + ; scale bar, 100 µm. (C) Representative immunoblots and quantification of p62 and Nrf2 levels in nrf2 +/+ , nrf2 -/- [Vector], nrf2 -/- [Flag-Nrf2] and nrf2 -/- [Flag-p62]. Data are presented as the mean ± SEM representative of three independent experiments and were assessed using one-way ANOVA followed by Tukey's test. * P

    Article Snippet: The following primary antibodies were used: Anti-Nrf2 (cat. no. ab137550; 1:2,000; Abcam), anti-p62 (cat.no.

    Techniques: Expressing, Knock-Out, Plasmid Preparation, Staining, Western Blot

    MAPK/p38 inhibitor attenuates aggresome formation through Nrf2-mediated transcriptional activation. (A) Representative images and quantification of aggresomes in DMSO, MG132 or MG132+TAK715 treated cells (12 h). Anti-UbK48 was used to visualize aggresomes (arrowheads) and DAPI was used to detect nuclei; scale bar, 20 µm. (B) Representative immunoblots and quantification of Nrf2 and p62 in treated cells. Data are presented as the mean ± SEM representative of three independent experiments and were assessed using one-way ANOVA followed by Tukey's test. * P

    Journal: Biomedical Reports

    Article Title: Transcriptional factor Nrf2 is essential for aggresome formation during proteasome inhibition

    doi: 10.3892/br.2019.1247

    Figure Lengend Snippet: MAPK/p38 inhibitor attenuates aggresome formation through Nrf2-mediated transcriptional activation. (A) Representative images and quantification of aggresomes in DMSO, MG132 or MG132+TAK715 treated cells (12 h). Anti-UbK48 was used to visualize aggresomes (arrowheads) and DAPI was used to detect nuclei; scale bar, 20 µm. (B) Representative immunoblots and quantification of Nrf2 and p62 in treated cells. Data are presented as the mean ± SEM representative of three independent experiments and were assessed using one-way ANOVA followed by Tukey's test. * P

    Article Snippet: The following primary antibodies were used: Anti-Nrf2 (cat. no. ab137550; 1:2,000; Abcam), anti-p62 (cat.no.

    Techniques: Activation Assay, Western Blot

    Transcriptional upregulation of p62 during proteasome inhibition is dependent on Nrf2. (A) Representative images and quantification of p62 mRNA in nrf2 +/+ and nrf2 -/- cells treated with DMSO or MG132 for 12 h. (B) Representative immunoblots and quantification of p62 levels in nrf2 +/+ and nrf2 -/- cells treated with DMSO or MG132 (12 h). (C) Representative images and quantification of EGFP expression driven by the p62 promoter in nrf2 +/+ and nrf2 -/- cells treated with DMSO or MG132 (12 h). Data are presented as the mean ± SEM representative of three independent experiments and were assessed using one-way ANOVA followed by Tukey's test. * P

    Journal: Biomedical Reports

    Article Title: Transcriptional factor Nrf2 is essential for aggresome formation during proteasome inhibition

    doi: 10.3892/br.2019.1247

    Figure Lengend Snippet: Transcriptional upregulation of p62 during proteasome inhibition is dependent on Nrf2. (A) Representative images and quantification of p62 mRNA in nrf2 +/+ and nrf2 -/- cells treated with DMSO or MG132 for 12 h. (B) Representative immunoblots and quantification of p62 levels in nrf2 +/+ and nrf2 -/- cells treated with DMSO or MG132 (12 h). (C) Representative images and quantification of EGFP expression driven by the p62 promoter in nrf2 +/+ and nrf2 -/- cells treated with DMSO or MG132 (12 h). Data are presented as the mean ± SEM representative of three independent experiments and were assessed using one-way ANOVA followed by Tukey's test. * P

    Article Snippet: The following primary antibodies were used: Anti-Nrf2 (cat. no. ab137550; 1:2,000; Abcam), anti-p62 (cat.no.

    Techniques: Inhibition, Western Blot, Expressing

    Loss of Nrf2 results in failure of proteasome inhibition-induced aggresome formation. (A) Representative images and quantification of aggresomes in nrf2 +/+ and nrf2 -/- cells after 14 h treatment with MG132 (2 µM). Anti-UbK48 was used to visualize aggresomes (arrowheads) and DAPI staining was used to detect nuclei; scale bar, 20 µm. (B) Representative images and quantification of nrf2 +/+ and nrf2 -/- cells stained with Annexin V (green) and PI (red) after 20 h treatment with DMSO or MG132 (2 µM). Live cells, Annexin V - PI - ; early apoptotic cells, Annexin V + PI - ; necrotic cells, Annexin V - PI + ; late apoptotic cells, Annexin V + PI + ; scale bar, 100 µm. Data are presented as the mean ± SEM representative of three independent experiments and were assessed using one-way ANOVA followed by Tukey's test. * P

    Journal: Biomedical Reports

    Article Title: Transcriptional factor Nrf2 is essential for aggresome formation during proteasome inhibition

    doi: 10.3892/br.2019.1247

    Figure Lengend Snippet: Loss of Nrf2 results in failure of proteasome inhibition-induced aggresome formation. (A) Representative images and quantification of aggresomes in nrf2 +/+ and nrf2 -/- cells after 14 h treatment with MG132 (2 µM). Anti-UbK48 was used to visualize aggresomes (arrowheads) and DAPI staining was used to detect nuclei; scale bar, 20 µm. (B) Representative images and quantification of nrf2 +/+ and nrf2 -/- cells stained with Annexin V (green) and PI (red) after 20 h treatment with DMSO or MG132 (2 µM). Live cells, Annexin V - PI - ; early apoptotic cells, Annexin V + PI - ; necrotic cells, Annexin V - PI + ; late apoptotic cells, Annexin V + PI + ; scale bar, 100 µm. Data are presented as the mean ± SEM representative of three independent experiments and were assessed using one-way ANOVA followed by Tukey's test. * P

    Article Snippet: The following primary antibodies were used: Anti-Nrf2 (cat. no. ab137550; 1:2,000; Abcam), anti-p62 (cat.no.

    Techniques: Inhibition, Staining

    Inhibition of protein synthesis blocks proteasome inhibitor-induced aggresome formation. (A) Representative images and quantification of aggresomes in DMSO, MG132, MG132+ANI and MG132+ActD treated cells. 293 cells were treated with ANI (5 µg/ml), ActD (2 µg/ml) and/or MG132 (2 µM) for 12 h and stained with anti-UbK48 to visualize aggresomes (arrowheads); scale bar, 20 µm. (B) Representative images and quantification of aggresomes in cells treated with MG132 (2 µM) and/or ANI (5 µg/ml) for the indicated periods and aggresomes were stained with anti-UbK48 (arrowheads); scale bar, 20 µm. (C) Representative immunoblots and quantification of Nrf2 and p62 in cells treated by MG132 (2 µM) and/or ANI (5 µg/ml). (D) Representative images and quantification of Nrf2 in DMSO and MG132 treated cells. The 293 cells were treated with MG132 (2 µM) for 12 h and stained with anti-Nrf2 antibody to visualize the localization of endogenous Nrf2; DAPI was used to detect nuclei; scale bar, 20 µm. Data are presented as the mean ± SEM and were assessed using one-way ANOVA followed by Tukey's test. *** P

    Journal: Biomedical Reports

    Article Title: Transcriptional factor Nrf2 is essential for aggresome formation during proteasome inhibition

    doi: 10.3892/br.2019.1247

    Figure Lengend Snippet: Inhibition of protein synthesis blocks proteasome inhibitor-induced aggresome formation. (A) Representative images and quantification of aggresomes in DMSO, MG132, MG132+ANI and MG132+ActD treated cells. 293 cells were treated with ANI (5 µg/ml), ActD (2 µg/ml) and/or MG132 (2 µM) for 12 h and stained with anti-UbK48 to visualize aggresomes (arrowheads); scale bar, 20 µm. (B) Representative images and quantification of aggresomes in cells treated with MG132 (2 µM) and/or ANI (5 µg/ml) for the indicated periods and aggresomes were stained with anti-UbK48 (arrowheads); scale bar, 20 µm. (C) Representative immunoblots and quantification of Nrf2 and p62 in cells treated by MG132 (2 µM) and/or ANI (5 µg/ml). (D) Representative images and quantification of Nrf2 in DMSO and MG132 treated cells. The 293 cells were treated with MG132 (2 µM) for 12 h and stained with anti-Nrf2 antibody to visualize the localization of endogenous Nrf2; DAPI was used to detect nuclei; scale bar, 20 µm. Data are presented as the mean ± SEM and were assessed using one-way ANOVA followed by Tukey's test. *** P

    Article Snippet: The following primary antibodies were used: Anti-Nrf2 (cat. no. ab137550; 1:2,000; Abcam), anti-p62 (cat.no.

    Techniques: Inhibition, Staining, Western Blot

    Pre-treatment with tert-butylhydroquinone (tBHQ) restores the total nuclear factor erythroid 2-related factor 2 (Nrf2) protein level and nuclear accumulation which are inhibited by ethanol (EtOH) in H9c2 cells. Western blot analysis was performed to analyze the level of Nrf2 protein. Cell lysates were prepared from H9c2 cells cultured in control medium (control), exposed to 200 mM EtOH or 5 µ M tBHQ (tBHQ) alone, or exposed to EtOH and pre-treated with tBHQ (EtOH + tBHQ). The data are expressed as fold change over control and represent the means ± SD of 3 separate experiments; * p

    Journal: International Journal of Molecular Medicine

    Article Title: Tert-butylhydroquinone attenuates the ethanol-induced apoptosis of and activates the Nrf2 antioxidant defense pathway in H9c2 cardiomyocytes

    doi: 10.3892/ijmm.2016.2605

    Figure Lengend Snippet: Pre-treatment with tert-butylhydroquinone (tBHQ) restores the total nuclear factor erythroid 2-related factor 2 (Nrf2) protein level and nuclear accumulation which are inhibited by ethanol (EtOH) in H9c2 cells. Western blot analysis was performed to analyze the level of Nrf2 protein. Cell lysates were prepared from H9c2 cells cultured in control medium (control), exposed to 200 mM EtOH or 5 µ M tBHQ (tBHQ) alone, or exposed to EtOH and pre-treated with tBHQ (EtOH + tBHQ). The data are expressed as fold change over control and represent the means ± SD of 3 separate experiments; * p

    Article Snippet: The cells were then incubated overnight with primary antibodies against AC-tubulin (6-11B-1; ab24610) at 1:200 dilution and Nrf2 (both from Abcam) at a 1:200 dilution in a humidified chamber at 4°C.

    Techniques: Western Blot, Cell Culture

    The immunofluorescence assay evidenced that tert-butylhydroquinone (tBHQ) promotes the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) in H9C2 cells. Nrf2 protein was visualized with a TRITC-labeled antibody, AC-tubulin was visualized with an FITC-labeled antibody, and the nuclear morphology was visualized with DAPI dye.

    Journal: International Journal of Molecular Medicine

    Article Title: Tert-butylhydroquinone attenuates the ethanol-induced apoptosis of and activates the Nrf2 antioxidant defense pathway in H9c2 cardiomyocytes

    doi: 10.3892/ijmm.2016.2605

    Figure Lengend Snippet: The immunofluorescence assay evidenced that tert-butylhydroquinone (tBHQ) promotes the nuclear translocation of nuclear factor erythroid 2-related factor 2 (Nrf2) in H9C2 cells. Nrf2 protein was visualized with a TRITC-labeled antibody, AC-tubulin was visualized with an FITC-labeled antibody, and the nuclear morphology was visualized with DAPI dye.

    Article Snippet: The cells were then incubated overnight with primary antibodies against AC-tubulin (6-11B-1; ab24610) at 1:200 dilution and Nrf2 (both from Abcam) at a 1:200 dilution in a humidified chamber at 4°C.

    Techniques: Immunofluorescence, Translocation Assay, Labeling

    Apoe deficiency increases oxidative stress after SCI. (A) Representative WBs of antioxidative proteins Nrf2, HO-1, NQO1 and the loading control (GAPDH) in WT and Apoe KO groups at 7 days after SCI. (B–D) Quantitative analysis of Nrf2, HO-1 and NQO1 expression. Data are expressed as mean ± SEM of three independent experiments ( n = 6 per group). * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Apolipoprotein E Deficiency Exacerbates Spinal Cord Injury in Mice: Inflammatory Response and Oxidative Stress Mediated by NF-κB Signaling Pathway

    doi: 10.3389/fncel.2018.00142

    Figure Lengend Snippet: Apoe deficiency increases oxidative stress after SCI. (A) Representative WBs of antioxidative proteins Nrf2, HO-1, NQO1 and the loading control (GAPDH) in WT and Apoe KO groups at 7 days after SCI. (B–D) Quantitative analysis of Nrf2, HO-1 and NQO1 expression. Data are expressed as mean ± SEM of three independent experiments ( n = 6 per group). * p

    Article Snippet: The membranes were blocked with 5% dried skimmed milk in TBS with 0.1% Tween-20 for 2 h at RT, and then incubated at 4°C overnight with the following primary antibodies: mouse anti-GAPDH (1:10,000, HC301-01, Transgene, Beijing, China), rabbit anti-APOE (1:200, sc-13520, Santa Cruz Biotechnology, CA, USA), mouse anti-GFAP (1:1000, ab7260, Abcam, Cambridge, England), mouse anti-IL-6 (1:1000, ab9324, Abcam, Cambridge, England), rabbit anti-IL-1β (1:1000, ab9722, Abcam Cambridge, England), rabbit anti-Bcl-2 (1:1000, ab136285, Abcam, Cambridge, England), rabbit anti-Bax (1:1000, ab32503, Abcam, Cambridge, England), rabbit anti-Nrf2 (1:1000, ab31163, Abcam, Cambridge, England), rabbit anti-p-NF-κB-p65 (1:1000, ab86299, Abcam, Cambridge, England), mouse anti-HO-1 (1:1000, ab3248, Abcam, Cambridge, England), mouse anti-NQO-1(1:1000, ab2947, Abcam, Cambridge, England), mouse anti-Erk1(pT202/pY204) + Erk2(Pt185/Py187; 1:1000, 50011, Abcam, Cambridge, England), rabbit anti-ERK1 + ERK2 (1:1000, 184699, Abcam, Cambridge, England), rabbit anti-Phosphp-p38-MAPK(Thr180/Tyr182; 1:1000, 9211, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-p38-MAPK (1:1000, 9212, Cell Signaling Technology, Danvers, MA, USA), and rabbit anti-JNK1 (pThr183/pThy185; 1:1000, NBP1-72242, Novus Biologicals, Littleton, CO, USA).

    Techniques: Expressing

    Apoe deficiency exacerbates inflammation by activating of NF-κB and inactivating of Nrf2 signaling pathways following SCI. (A) Representative immunoblots for p-NF–κB, IL-6, IL-1β and the loading control (GAPDH) in WT and Apoe KO groups at 7 days after SCI. (B–D) Quantitative levels of p-NF-κB, IL-6 and IL-1β in lesion area of Apoe KO and WT mice at 7 days post-injury. Data are expressed as mean ± SEM of three independent experiments ( n = 6 per group). * p

    Journal: Frontiers in Cellular Neuroscience

    Article Title: Apolipoprotein E Deficiency Exacerbates Spinal Cord Injury in Mice: Inflammatory Response and Oxidative Stress Mediated by NF-κB Signaling Pathway

    doi: 10.3389/fncel.2018.00142

    Figure Lengend Snippet: Apoe deficiency exacerbates inflammation by activating of NF-κB and inactivating of Nrf2 signaling pathways following SCI. (A) Representative immunoblots for p-NF–κB, IL-6, IL-1β and the loading control (GAPDH) in WT and Apoe KO groups at 7 days after SCI. (B–D) Quantitative levels of p-NF-κB, IL-6 and IL-1β in lesion area of Apoe KO and WT mice at 7 days post-injury. Data are expressed as mean ± SEM of three independent experiments ( n = 6 per group). * p

    Article Snippet: The membranes were blocked with 5% dried skimmed milk in TBS with 0.1% Tween-20 for 2 h at RT, and then incubated at 4°C overnight with the following primary antibodies: mouse anti-GAPDH (1:10,000, HC301-01, Transgene, Beijing, China), rabbit anti-APOE (1:200, sc-13520, Santa Cruz Biotechnology, CA, USA), mouse anti-GFAP (1:1000, ab7260, Abcam, Cambridge, England), mouse anti-IL-6 (1:1000, ab9324, Abcam, Cambridge, England), rabbit anti-IL-1β (1:1000, ab9722, Abcam Cambridge, England), rabbit anti-Bcl-2 (1:1000, ab136285, Abcam, Cambridge, England), rabbit anti-Bax (1:1000, ab32503, Abcam, Cambridge, England), rabbit anti-Nrf2 (1:1000, ab31163, Abcam, Cambridge, England), rabbit anti-p-NF-κB-p65 (1:1000, ab86299, Abcam, Cambridge, England), mouse anti-HO-1 (1:1000, ab3248, Abcam, Cambridge, England), mouse anti-NQO-1(1:1000, ab2947, Abcam, Cambridge, England), mouse anti-Erk1(pT202/pY204) + Erk2(Pt185/Py187; 1:1000, 50011, Abcam, Cambridge, England), rabbit anti-ERK1 + ERK2 (1:1000, 184699, Abcam, Cambridge, England), rabbit anti-Phosphp-p38-MAPK(Thr180/Tyr182; 1:1000, 9211, Cell Signaling Technology, Danvers, MA, USA), rabbit anti-p38-MAPK (1:1000, 9212, Cell Signaling Technology, Danvers, MA, USA), and rabbit anti-JNK1 (pThr183/pThy185; 1:1000, NBP1-72242, Novus Biologicals, Littleton, CO, USA).

    Techniques: Western Blot, Mouse Assay

    Protective effect of eriodictyol-induced HO-1 expression on H 2 O 2 -induced cell damage. ( A ) H 2 O 2 -exposed cells were pretreated for 1 h with or without ZnPP and then treated with eriodictyol. The inhibitory effect of eriodictyol on H 2 O 2 -induced intracellular ROS generation was observed by fluorescence microscopy; ( B ) H 2 O 2 -stimulated cells were pretreated for 1 h with or without ZnPP or Nrf2 or HO-1 siRNA and then treated with eriodictyol. Protective effect of HO-1 induction on cell death was determined by in situ terminal nick end-labeling (TUNEL) assay: −, untreated; +, treated. ROS and TUNEL staining was quantified in four randomly selected fields for each group: C, control. The scale bars for image ( B ) are the same as that in ( A ), 100 μm. * p

    Journal: International Journal of Molecular Sciences

    Article Title: Eriodictyol Protects Endothelial Cells against Oxidative Stress-Induced Cell Death through Modulating ERK/Nrf2/ARE-Dependent Heme Oxygenase-1 Expression

    doi: 10.3390/ijms160714526

    Figure Lengend Snippet: Protective effect of eriodictyol-induced HO-1 expression on H 2 O 2 -induced cell damage. ( A ) H 2 O 2 -exposed cells were pretreated for 1 h with or without ZnPP and then treated with eriodictyol. The inhibitory effect of eriodictyol on H 2 O 2 -induced intracellular ROS generation was observed by fluorescence microscopy; ( B ) H 2 O 2 -stimulated cells were pretreated for 1 h with or without ZnPP or Nrf2 or HO-1 siRNA and then treated with eriodictyol. Protective effect of HO-1 induction on cell death was determined by in situ terminal nick end-labeling (TUNEL) assay: −, untreated; +, treated. ROS and TUNEL staining was quantified in four randomly selected fields for each group: C, control. The scale bars for image ( B ) are the same as that in ( A ), 100 μm. * p

    Article Snippet: Western Blot Analysis After eriodictyol treatment, cells were washed with phosphate-buffered saline and mixed with RIPA (50 mM Tris CL, pH 7.4/150 mM NaCl/1% Nonidet P-40 (NP-40)/1% sodium deoxycholate/0.1% SDS) buffer containing 1 mM etilendiaminetetraacetic acid (EDTA), 5 µg/mL aprotinin, 2 µg/mL leupeptin, and 1 mM phenylmethylsulfonyl fluoride (PMSF), followed by centrifugation at 14,000× g for 15 min. We applied 20 μg of the whole cell lysate protein to each lane and analyzed them by Western blot, using a monoclonal antibody against HO-1, anti-Nrf2, anti-Lamin B, and GAPDH).

    Techniques: Expressing, Fluorescence, Microscopy, In Situ, End Labeling, TUNEL Assay, Staining

    Nrf2 nuclear translocation induced by eriodictyol. ( A ) Cells were treated with 10 and 20 μM eriodictyol at the indicated concentrations for 4 h. Nuclear extracts were subjected to Western blot, using an anti-Nrf2 antibody and anti-lamin B antibody (a marker of nuclear protein); and ( B ) Transient transfection of cells with increasing doses of Nrf2-specific siRNA (10 and 20 nM) reduced HO-1 expression. Western blots representative of three independent experiments are shown: C, untreated cells; +, eriodictyol treatment only; , dose increasing.

    Journal: International Journal of Molecular Sciences

    Article Title: Eriodictyol Protects Endothelial Cells against Oxidative Stress-Induced Cell Death through Modulating ERK/Nrf2/ARE-Dependent Heme Oxygenase-1 Expression

    doi: 10.3390/ijms160714526

    Figure Lengend Snippet: Nrf2 nuclear translocation induced by eriodictyol. ( A ) Cells were treated with 10 and 20 μM eriodictyol at the indicated concentrations for 4 h. Nuclear extracts were subjected to Western blot, using an anti-Nrf2 antibody and anti-lamin B antibody (a marker of nuclear protein); and ( B ) Transient transfection of cells with increasing doses of Nrf2-specific siRNA (10 and 20 nM) reduced HO-1 expression. Western blots representative of three independent experiments are shown: C, untreated cells; +, eriodictyol treatment only; , dose increasing.

    Article Snippet: Western Blot Analysis After eriodictyol treatment, cells were washed with phosphate-buffered saline and mixed with RIPA (50 mM Tris CL, pH 7.4/150 mM NaCl/1% Nonidet P-40 (NP-40)/1% sodium deoxycholate/0.1% SDS) buffer containing 1 mM etilendiaminetetraacetic acid (EDTA), 5 µg/mL aprotinin, 2 µg/mL leupeptin, and 1 mM phenylmethylsulfonyl fluoride (PMSF), followed by centrifugation at 14,000× g for 15 min. We applied 20 μg of the whole cell lysate protein to each lane and analyzed them by Western blot, using a monoclonal antibody against HO-1, anti-Nrf2, anti-Lamin B, and GAPDH).

    Techniques: Translocation Assay, Western Blot, Marker, Transfection, Expressing

    NGR1 inhibits oxidative stress by up-regulating the Akt/Nrf2/HO-1 pathway in the hippocampus of db/db mice (A) SOD activity in the hippocampus of each group. (B) MDA levels in the hippocampus of each group. (C) Protein carbonyl levels in the hippocampus of each group. (D) Representative protein bands and Western blot analysis of Akt, p-Akt, Nrf2, HO-1, and TXNIP in the hippocampus of each group. Values are represented as means ± SD for 3 mice in each group. ## P

    Journal: Oncotarget

    Article Title: Notoginsenoside R1 ameliorates diabetic encephalopathy by activating the Nrf2 pathway and inhibiting NLRP3 inflammasome activation

    doi: 10.18632/oncotarget.24295

    Figure Lengend Snippet: NGR1 inhibits oxidative stress by up-regulating the Akt/Nrf2/HO-1 pathway in the hippocampus of db/db mice (A) SOD activity in the hippocampus of each group. (B) MDA levels in the hippocampus of each group. (C) Protein carbonyl levels in the hippocampus of each group. (D) Representative protein bands and Western blot analysis of Akt, p-Akt, Nrf2, HO-1, and TXNIP in the hippocampus of each group. Values are represented as means ± SD for 3 mice in each group. ## P

    Article Snippet: Primary antibodies against Akt, ASC, IL-1β, and Nrf2 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Mouse Assay, Activity Assay, Multiple Displacement Amplification, Western Blot

    NGR1 activates the Akt/Nrf2/HO-1 pathway, and inhibits NLRP3 inflammasome activation in HG-induced HT22 hippocampal neurons (A) Representative protein bands and Western blot analysis of NLRP3, ASC, and IL-1β in hippocampal neurons. (B) Caspase-1 activity in hippocampal neurons. (C) Representative protein bands and Western blot analysis of Akt, p-Akt, Nrf2, HO-1, and TXNIP in hippocampal neurons. Values are represented as means ± SD from three independent experiments. ## P

    Journal: Oncotarget

    Article Title: Notoginsenoside R1 ameliorates diabetic encephalopathy by activating the Nrf2 pathway and inhibiting NLRP3 inflammasome activation

    doi: 10.18632/oncotarget.24295

    Figure Lengend Snippet: NGR1 activates the Akt/Nrf2/HO-1 pathway, and inhibits NLRP3 inflammasome activation in HG-induced HT22 hippocampal neurons (A) Representative protein bands and Western blot analysis of NLRP3, ASC, and IL-1β in hippocampal neurons. (B) Caspase-1 activity in hippocampal neurons. (C) Representative protein bands and Western blot analysis of Akt, p-Akt, Nrf2, HO-1, and TXNIP in hippocampal neurons. Values are represented as means ± SD from three independent experiments. ## P

    Article Snippet: Primary antibodies against Akt, ASC, IL-1β, and Nrf2 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Activation Assay, Western Blot, Activity Assay

    NGR1 exerts neuroprotective effects and inhibition of NLRP3 inflammasome by activating the Akt/Nrf2 pathway (A) Cell viability was measured by MTT assay. (B) Caspase-3 activity in HT22 hippocampal neurons. (C) LDH release in HT22 hippocampal neurons. (D) Representative protein bands and Western blot analysis of NLRP3, ASC, and IL-1β in hippocampal neurons. (E) Caspase-1 activity in hippocampal neurons. (F) Representative protein bands and Western blot analysis of Akt, p-Akt, Nrf2, HO-1, and TXNIP in hippocampal neurons. Values are represented as means ± SD from three independent experiments. ## P

    Journal: Oncotarget

    Article Title: Notoginsenoside R1 ameliorates diabetic encephalopathy by activating the Nrf2 pathway and inhibiting NLRP3 inflammasome activation

    doi: 10.18632/oncotarget.24295

    Figure Lengend Snippet: NGR1 exerts neuroprotective effects and inhibition of NLRP3 inflammasome by activating the Akt/Nrf2 pathway (A) Cell viability was measured by MTT assay. (B) Caspase-3 activity in HT22 hippocampal neurons. (C) LDH release in HT22 hippocampal neurons. (D) Representative protein bands and Western blot analysis of NLRP3, ASC, and IL-1β in hippocampal neurons. (E) Caspase-1 activity in hippocampal neurons. (F) Representative protein bands and Western blot analysis of Akt, p-Akt, Nrf2, HO-1, and TXNIP in hippocampal neurons. Values are represented as means ± SD from three independent experiments. ## P

    Article Snippet: Primary antibodies against Akt, ASC, IL-1β, and Nrf2 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Inhibition, MTT Assay, Activity Assay, Western Blot

    Schematic of NGR1 mechanism of ameliorating DEP by activating the Akt/Nrf2/HO-1 pathway and inhibiting NLRP3 inflammasome activation

    Journal: Oncotarget

    Article Title: Notoginsenoside R1 ameliorates diabetic encephalopathy by activating the Nrf2 pathway and inhibiting NLRP3 inflammasome activation

    doi: 10.18632/oncotarget.24295

    Figure Lengend Snippet: Schematic of NGR1 mechanism of ameliorating DEP by activating the Akt/Nrf2/HO-1 pathway and inhibiting NLRP3 inflammasome activation

    Article Snippet: Primary antibodies against Akt, ASC, IL-1β, and Nrf2 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

    Techniques: Activation Assay

    Andrographolide induces Nrf2 expression in Ava5 cells. (A–C) Induction of Nrf2 expression (A), Nrf2 nuclear translocation (B) and Nrf2-mediated ARE transactivation (C) by andrographolide in a concentration- or time-dependent manner. Ava5 or p2xARE-Luc-transfected

    Journal: British Journal of Pharmacology

    Article Title: Andrographolide exerts anti-hepatitis C virus activity by up-regulating haeme oxygenase-1 via the p38 MAPK/Nrf2 pathway in human hepatoma cells

    doi: 10.1111/bph.12440

    Figure Lengend Snippet: Andrographolide induces Nrf2 expression in Ava5 cells. (A–C) Induction of Nrf2 expression (A), Nrf2 nuclear translocation (B) and Nrf2-mediated ARE transactivation (C) by andrographolide in a concentration- or time-dependent manner. Ava5 or p2xARE-Luc-transfected

    Article Snippet: The membranes were probed with anti-HCV NS5B (1:5000; Abcam, Cambridge, MA, USA), anti-HO-1 (1:3000; Abcam), anti-BVR antibody (1:1000; Abcam), anti-Nrf2 (1:3000; GeneTex, Irvine, CA, USA), anti-Keap1 (1:1000; GeneTex), anti-Bach1 (1:1000; Abcam), anti-phospho-ERK1/2 (1:1000; Cell Signaling Technology, Inc., Danvers, MA, USA), anti-phospho-p38 (1:1000; Cell Signaling), anti-phospho-JNK (1:1000; Cell Signaling), anti- ERK1/2 (1:1000; Cell Signaling), anti-p38 (1:1000; Cell Signaling), anti-JNK (1:1000; Cell Signaling) or anti-lamin B1 (1:1000; GeneTex) antibody.

    Techniques: Expressing, Translocation Assay, Concentration Assay, Transfection

    Andrographolide activates p38 MAPK phosphorylation to stimulate the Nrf2–HO-1 signalling pathway for anti-HCV activity. (A) Andrographolide activated p38 MAPK phosphorylation in a time-dependent manner. Ava5 cells were treated with 10 μM

    Journal: British Journal of Pharmacology

    Article Title: Andrographolide exerts anti-hepatitis C virus activity by up-regulating haeme oxygenase-1 via the p38 MAPK/Nrf2 pathway in human hepatoma cells

    doi: 10.1111/bph.12440

    Figure Lengend Snippet: Andrographolide activates p38 MAPK phosphorylation to stimulate the Nrf2–HO-1 signalling pathway for anti-HCV activity. (A) Andrographolide activated p38 MAPK phosphorylation in a time-dependent manner. Ava5 cells were treated with 10 μM

    Article Snippet: The membranes were probed with anti-HCV NS5B (1:5000; Abcam, Cambridge, MA, USA), anti-HO-1 (1:3000; Abcam), anti-BVR antibody (1:1000; Abcam), anti-Nrf2 (1:3000; GeneTex, Irvine, CA, USA), anti-Keap1 (1:1000; GeneTex), anti-Bach1 (1:1000; Abcam), anti-phospho-ERK1/2 (1:1000; Cell Signaling Technology, Inc., Danvers, MA, USA), anti-phospho-p38 (1:1000; Cell Signaling), anti-phospho-JNK (1:1000; Cell Signaling), anti- ERK1/2 (1:1000; Cell Signaling), anti-p38 (1:1000; Cell Signaling), anti-JNK (1:1000; Cell Signaling) or anti-lamin B1 (1:1000; GeneTex) antibody.

    Techniques: Activity Assay

    Model describing the inhibition of HCV replication by andrographolide. Andrographolide induces p38 MAPK activation and subsequently increases Nrf2-mediated HO-1 expression, leading to the generation of biliverdin against HCV replication by enhancement

    Journal: British Journal of Pharmacology

    Article Title: Andrographolide exerts anti-hepatitis C virus activity by up-regulating haeme oxygenase-1 via the p38 MAPK/Nrf2 pathway in human hepatoma cells

    doi: 10.1111/bph.12440

    Figure Lengend Snippet: Model describing the inhibition of HCV replication by andrographolide. Andrographolide induces p38 MAPK activation and subsequently increases Nrf2-mediated HO-1 expression, leading to the generation of biliverdin against HCV replication by enhancement

    Article Snippet: The membranes were probed with anti-HCV NS5B (1:5000; Abcam, Cambridge, MA, USA), anti-HO-1 (1:3000; Abcam), anti-BVR antibody (1:1000; Abcam), anti-Nrf2 (1:3000; GeneTex, Irvine, CA, USA), anti-Keap1 (1:1000; GeneTex), anti-Bach1 (1:1000; Abcam), anti-phospho-ERK1/2 (1:1000; Cell Signaling Technology, Inc., Danvers, MA, USA), anti-phospho-p38 (1:1000; Cell Signaling), anti-phospho-JNK (1:1000; Cell Signaling), anti- ERK1/2 (1:1000; Cell Signaling), anti-p38 (1:1000; Cell Signaling), anti-JNK (1:1000; Cell Signaling) or anti-lamin B1 (1:1000; GeneTex) antibody.

    Techniques: Inhibition, Activation Assay, Expressing

    Scheme of SFN-mediated inhibition of HCV replication. SFN induces PI3K phosphorylation and subsequently triggers Nrf2 nuclear translocation to activate HO-1 expression leading to biliverdin production against HCV replication by the induction of antiviral IFN responses and suppression of HCV NS3/4A protease activity.

    Journal: PLoS ONE

    Article Title: Sulforaphane Suppresses Hepatitis C Virus Replication by Up-Regulating Heme Oxygenase-1 Expression through PI3K/Nrf2 Pathway

    doi: 10.1371/journal.pone.0152236

    Figure Lengend Snippet: Scheme of SFN-mediated inhibition of HCV replication. SFN induces PI3K phosphorylation and subsequently triggers Nrf2 nuclear translocation to activate HO-1 expression leading to biliverdin production against HCV replication by the induction of antiviral IFN responses and suppression of HCV NS3/4A protease activity.

    Article Snippet: The membranes were blocked with 5% nonfat milk in phosphate-buffered saline containing Tween 20 (PBST) and incubated overnight at 4°C with primary antibodies against GAPDH (1:10,000; GeneTex, CA, USA), NS5B (1:5,000; Abcam, Cambridge, MA, USA), Nrf2 (1:3,000; GeneTex), HO-1 (1:3,000; GeneTex), biliverdin reductase A (BVRA), Keap1 and Bach1 (1:1,000; Abcam), and total-phohphoinositide 3-kinase (PI3K) or phosphor-PI3K (1:1000; Cell Signaling Technology, Inc., Beverly, MA, USA).

    Techniques: Inhibition, Translocation Assay, Expressing, Activity Assay

    SFN inhibited HCV replication by upregulating Nrf2 expression. SFN stimulated (A) ARE transactivation, (B) Nrf2 expression, and (C) Nrf2 nuclear translocation in a concentration- or time-dependent manner. The antioxidant response reporter plasmid, p3xARE-Luc, was transfected into Ava5 cells and then treated with the indicated concentrations of SFN (0–10 μM) for 3 days. The relative induction of antioxidant activity was determined by luciferase assay. The activity of SFN-untreated Ava5 cells was considered to be 1. The lysates of cytoplasmic and nuclear fractions were separated and the expression levels of HO-1 regulators were analyzed by Western blotting. The SFN-induced Nrf2 nuclear translocation was analyzed by Western blotting at different incubation times (0–72 h). (D and E) Anti-HCV activity of SFN was attenuated by Nrf2 shRNA against Nrf2 expression. Increasing amounts of Nrf2-specific shRNA (0.25–2 μg) or non-specific shRNA were transfected into Ava5 cells. The transfected cells were then treated with 7.5 μM SFN for 3 days. HCV and HO-1 protein and RNA levels were analyzed by Western blotting and qRT-PCR, respectively. Western blotting was performed using antibodies against HCV NS5B, Nrf2, Keap1, Bach1, and HO-1. The expression levels of GAPDH and Histone 1 showed equal loading of cell lysates. Relative RNA levels were normalized to the internal control gapdh . The relative HCV RNA levels were presented as percentage changes compared to SFN-untreated/untransfected Ava5 cells, in which the level was considered to be 100%. Data were represented as the means of normalized data ± standard deviations (error bars) based on five independent experiments. * P

    Journal: PLoS ONE

    Article Title: Sulforaphane Suppresses Hepatitis C Virus Replication by Up-Regulating Heme Oxygenase-1 Expression through PI3K/Nrf2 Pathway

    doi: 10.1371/journal.pone.0152236

    Figure Lengend Snippet: SFN inhibited HCV replication by upregulating Nrf2 expression. SFN stimulated (A) ARE transactivation, (B) Nrf2 expression, and (C) Nrf2 nuclear translocation in a concentration- or time-dependent manner. The antioxidant response reporter plasmid, p3xARE-Luc, was transfected into Ava5 cells and then treated with the indicated concentrations of SFN (0–10 μM) for 3 days. The relative induction of antioxidant activity was determined by luciferase assay. The activity of SFN-untreated Ava5 cells was considered to be 1. The lysates of cytoplasmic and nuclear fractions were separated and the expression levels of HO-1 regulators were analyzed by Western blotting. The SFN-induced Nrf2 nuclear translocation was analyzed by Western blotting at different incubation times (0–72 h). (D and E) Anti-HCV activity of SFN was attenuated by Nrf2 shRNA against Nrf2 expression. Increasing amounts of Nrf2-specific shRNA (0.25–2 μg) or non-specific shRNA were transfected into Ava5 cells. The transfected cells were then treated with 7.5 μM SFN for 3 days. HCV and HO-1 protein and RNA levels were analyzed by Western blotting and qRT-PCR, respectively. Western blotting was performed using antibodies against HCV NS5B, Nrf2, Keap1, Bach1, and HO-1. The expression levels of GAPDH and Histone 1 showed equal loading of cell lysates. Relative RNA levels were normalized to the internal control gapdh . The relative HCV RNA levels were presented as percentage changes compared to SFN-untreated/untransfected Ava5 cells, in which the level was considered to be 100%. Data were represented as the means of normalized data ± standard deviations (error bars) based on five independent experiments. * P

    Article Snippet: The membranes were blocked with 5% nonfat milk in phosphate-buffered saline containing Tween 20 (PBST) and incubated overnight at 4°C with primary antibodies against GAPDH (1:10,000; GeneTex, CA, USA), NS5B (1:5,000; Abcam, Cambridge, MA, USA), Nrf2 (1:3,000; GeneTex), HO-1 (1:3,000; GeneTex), biliverdin reductase A (BVRA), Keap1 and Bach1 (1:1,000; Abcam), and total-phohphoinositide 3-kinase (PI3K) or phosphor-PI3K (1:1000; Cell Signaling Technology, Inc., Beverly, MA, USA).

    Techniques: Expressing, Translocation Assay, Concentration Assay, Plasmid Preparation, Transfection, Antioxidant Activity Assay, Luciferase, Activity Assay, Western Blot, Incubation, shRNA, Quantitative RT-PCR

    SFN-mediated Nrf2/HO-1 activation was PI3K dependent. (A) Phosphorylation of PI3K was induced by SFN treatment. Ava5 cells were treated with 7.5 μM SFN, the cellular lysates were collected at the indicated time points (0–120 min), and the phosphorylation level of PI3K was analyzed by Western blotting. (B) Anti-HCV activity of SFN was attenuated by PI3K inhibition. Ava5 cells were treated with 7.5 μM SFN with or without the PI3K-specific inhibitor, wortmanin (0–10 μM), for 3 days. The HCV protein, Nrf2, and HO-1 expression were analyzed by Western blotting. Western blotting that was performed using antibodies against phospho-PI3K, total-PI3K, HCV NS5B, Nrf2, and HO-1. An antibody against GAPDH was used to show equal loading of lysates.

    Journal: PLoS ONE

    Article Title: Sulforaphane Suppresses Hepatitis C Virus Replication by Up-Regulating Heme Oxygenase-1 Expression through PI3K/Nrf2 Pathway

    doi: 10.1371/journal.pone.0152236

    Figure Lengend Snippet: SFN-mediated Nrf2/HO-1 activation was PI3K dependent. (A) Phosphorylation of PI3K was induced by SFN treatment. Ava5 cells were treated with 7.5 μM SFN, the cellular lysates were collected at the indicated time points (0–120 min), and the phosphorylation level of PI3K was analyzed by Western blotting. (B) Anti-HCV activity of SFN was attenuated by PI3K inhibition. Ava5 cells were treated with 7.5 μM SFN with or without the PI3K-specific inhibitor, wortmanin (0–10 μM), for 3 days. The HCV protein, Nrf2, and HO-1 expression were analyzed by Western blotting. Western blotting that was performed using antibodies against phospho-PI3K, total-PI3K, HCV NS5B, Nrf2, and HO-1. An antibody against GAPDH was used to show equal loading of lysates.

    Article Snippet: The membranes were blocked with 5% nonfat milk in phosphate-buffered saline containing Tween 20 (PBST) and incubated overnight at 4°C with primary antibodies against GAPDH (1:10,000; GeneTex, CA, USA), NS5B (1:5,000; Abcam, Cambridge, MA, USA), Nrf2 (1:3,000; GeneTex), HO-1 (1:3,000; GeneTex), biliverdin reductase A (BVRA), Keap1 and Bach1 (1:1,000; Abcam), and total-phohphoinositide 3-kinase (PI3K) or phosphor-PI3K (1:1000; Cell Signaling Technology, Inc., Beverly, MA, USA).

    Techniques: Activation Assay, Western Blot, Activity Assay, Inhibition, Expressing

    DJ-1 does not impact the interaction between Nrf2 and Keap1. ( A ) DJ-1 does not interact with Nrf2. HeLa cells were transfected with Flag-Nrf2 and Myc-DJ-1 for 48 h. Cells were washed twice with PBS, and dithiobis[succinimidylpropionate] (DSP, Thermo Scientific)

    Journal: Human Molecular Genetics

    Article Title: DJ-1 induces thioredoxin 1 expression through the Nrf2 pathway

    doi: 10.1093/hmg/dds131

    Figure Lengend Snippet: DJ-1 does not impact the interaction between Nrf2 and Keap1. ( A ) DJ-1 does not interact with Nrf2. HeLa cells were transfected with Flag-Nrf2 and Myc-DJ-1 for 48 h. Cells were washed twice with PBS, and dithiobis[succinimidylpropionate] (DSP, Thermo Scientific)

    Article Snippet: Antibodies used for western blot analyses and their sources are as follows: anti-Trx1, peroxidase-conjugated anti-Myc, anti-Flag and anti-Lamin B1 from Santa Cruz Biotechnology; DJ-1 (691, a kind gift from Benoit I. Giasson) and anti-human DJ-1 from Stressgen; anti-Nrf2 from Epitomics; anti-pAKT (S473) and anti-AKT from Cell Signaling; anti-β-actin and anti-α-tubulin from Sigma.

    Techniques: Transfection

    Nrf2 mediates DJ-1-dependent Trx1 expression and cytoprotection. ( A ) Nrf2 knockdown reduces Trx1 expression. Flag-DJ-1 and empty-vector-engineered cells were transfected with siRNA-targeting Nrf2 (siNrf2), and western blots were performed 72 h later.

    Journal: Human Molecular Genetics

    Article Title: DJ-1 induces thioredoxin 1 expression through the Nrf2 pathway

    doi: 10.1093/hmg/dds131

    Figure Lengend Snippet: Nrf2 mediates DJ-1-dependent Trx1 expression and cytoprotection. ( A ) Nrf2 knockdown reduces Trx1 expression. Flag-DJ-1 and empty-vector-engineered cells were transfected with siRNA-targeting Nrf2 (siNrf2), and western blots were performed 72 h later.

    Article Snippet: Antibodies used for western blot analyses and their sources are as follows: anti-Trx1, peroxidase-conjugated anti-Myc, anti-Flag and anti-Lamin B1 from Santa Cruz Biotechnology; DJ-1 (691, a kind gift from Benoit I. Giasson) and anti-human DJ-1 from Stressgen; anti-Nrf2 from Epitomics; anti-pAKT (S473) and anti-AKT from Cell Signaling; anti-β-actin and anti-α-tubulin from Sigma.

    Techniques: Expressing, Plasmid Preparation, Transfection, Western Blot

    DJ-1 stimulates nuclear translocation of Nrf2 and enhances its recruitment to the Trx1 promoter. ( A ) Overexpression of DJ-1 enhances nuclear translocation of Nrf2. SY5Y (−) and SY5Y-DJ-1 (+) cell lysates were fractionated to nuclear and cytoplasmic

    Journal: Human Molecular Genetics

    Article Title: DJ-1 induces thioredoxin 1 expression through the Nrf2 pathway

    doi: 10.1093/hmg/dds131

    Figure Lengend Snippet: DJ-1 stimulates nuclear translocation of Nrf2 and enhances its recruitment to the Trx1 promoter. ( A ) Overexpression of DJ-1 enhances nuclear translocation of Nrf2. SY5Y (−) and SY5Y-DJ-1 (+) cell lysates were fractionated to nuclear and cytoplasmic

    Article Snippet: Antibodies used for western blot analyses and their sources are as follows: anti-Trx1, peroxidase-conjugated anti-Myc, anti-Flag and anti-Lamin B1 from Santa Cruz Biotechnology; DJ-1 (691, a kind gift from Benoit I. Giasson) and anti-human DJ-1 from Stressgen; anti-Nrf2 from Epitomics; anti-pAKT (S473) and anti-AKT from Cell Signaling; anti-β-actin and anti-α-tubulin from Sigma.

    Techniques: Translocation Assay, Over Expression

    DJ-1 increases Nrf2 protein levels. ( A ) No change of Nrf2 mRNA in the presence of DJ-1. Real-time PCR analyses of Nrf2 mRNA in SY5Y and SY5Y-DJ-1 cells. Relative mRNA levels were normalized to GAPDH levels. ( B ) Increase in the Nrf2 protein in the presence

    Journal: Human Molecular Genetics

    Article Title: DJ-1 induces thioredoxin 1 expression through the Nrf2 pathway

    doi: 10.1093/hmg/dds131

    Figure Lengend Snippet: DJ-1 increases Nrf2 protein levels. ( A ) No change of Nrf2 mRNA in the presence of DJ-1. Real-time PCR analyses of Nrf2 mRNA in SY5Y and SY5Y-DJ-1 cells. Relative mRNA levels were normalized to GAPDH levels. ( B ) Increase in the Nrf2 protein in the presence

    Article Snippet: Antibodies used for western blot analyses and their sources are as follows: anti-Trx1, peroxidase-conjugated anti-Myc, anti-Flag and anti-Lamin B1 from Santa Cruz Biotechnology; DJ-1 (691, a kind gift from Benoit I. Giasson) and anti-human DJ-1 from Stressgen; anti-Nrf2 from Epitomics; anti-pAKT (S473) and anti-AKT from Cell Signaling; anti-β-actin and anti-α-tubulin from Sigma.

    Techniques: Real-time Polymerase Chain Reaction

    (A) Representative immunoblots of select markers of Nrf2-mediated protective response. (B) Densitometric analyses of immunoblots.

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    Article Title: Pharmacologic Suppression of Inflammation by a Diphenyldifluoroketone, EF24, in a Rat Model of Fixed-Volume Hemorrhage Improves Survival

    doi: 10.1124/jpet.113.208009

    Figure Lengend Snippet: (A) Representative immunoblots of select markers of Nrf2-mediated protective response. (B) Densitometric analyses of immunoblots.

    Article Snippet: The proteins were fractionated by SDS-polyacrylamide gel electrophoresis, electrotransferred to the nitrocellulose membranes, blotted with primary antibodies against phospho-NF- κ B-p65 (3033; Cell Signaling Technology), TLR4 (2219; Cell Signaling Technology), COX-2 (sc1746; Santa Cruz Biotechnology), HMGB1 (3935; Cell Signaling Technology), inducible nitric-oxide synthase (iNOS) (ab15323; Abcam), heme oxygenase–1 (HO-1) (sc1797; Santa Cruz Biotechnology), nuclear factor (erythroid-derived 2)-like 2 (Nrf2) (2178-1; Epitomics), phospho-Nrf2 (2073-1; Epitomics), apurinic/apyrimidinic endonuclease (APE1) (4128; Cell Signaling Technology), superoxide dismutase–1 (SOD1) (2770; Cell Signaling Technology), and 8-oxoguanine glycosylase 1 (OGG1) (sc33181; Santa Cruz Biotech), followed by HRP-conjugated secondary antibody.

    Techniques: Western Blot

    The pleiotropic effects of EF24 against inflammation. The oxidative stress generated by blood loss recruits Nrf2 response on one hand and the TLR4/NF- κ B axis on the other hand. Both pathways are part of a protective response initiated by the hypoperfusion-induced tissue injury. Whereas the Nrf2-mediated antioxidant response in severe hemorrhage may not be sufficient to salvage the tissue, NF- κ B-driven inflammatory cascade may quickly attain exaggerated levels detrimental to the organism as a whole. The salutary effects of EF24 treatment in hemorrhage appear to be because it enhances the Nrf2-mediated antioxidant response and suppresses the NF- κ B-driven inflammation.

    Journal: The Journal of Pharmacology and Experimental Therapeutics

    Article Title: Pharmacologic Suppression of Inflammation by a Diphenyldifluoroketone, EF24, in a Rat Model of Fixed-Volume Hemorrhage Improves Survival

    doi: 10.1124/jpet.113.208009

    Figure Lengend Snippet: The pleiotropic effects of EF24 against inflammation. The oxidative stress generated by blood loss recruits Nrf2 response on one hand and the TLR4/NF- κ B axis on the other hand. Both pathways are part of a protective response initiated by the hypoperfusion-induced tissue injury. Whereas the Nrf2-mediated antioxidant response in severe hemorrhage may not be sufficient to salvage the tissue, NF- κ B-driven inflammatory cascade may quickly attain exaggerated levels detrimental to the organism as a whole. The salutary effects of EF24 treatment in hemorrhage appear to be because it enhances the Nrf2-mediated antioxidant response and suppresses the NF- κ B-driven inflammation.

    Article Snippet: The proteins were fractionated by SDS-polyacrylamide gel electrophoresis, electrotransferred to the nitrocellulose membranes, blotted with primary antibodies against phospho-NF- κ B-p65 (3033; Cell Signaling Technology), TLR4 (2219; Cell Signaling Technology), COX-2 (sc1746; Santa Cruz Biotechnology), HMGB1 (3935; Cell Signaling Technology), inducible nitric-oxide synthase (iNOS) (ab15323; Abcam), heme oxygenase–1 (HO-1) (sc1797; Santa Cruz Biotechnology), nuclear factor (erythroid-derived 2)-like 2 (Nrf2) (2178-1; Epitomics), phospho-Nrf2 (2073-1; Epitomics), apurinic/apyrimidinic endonuclease (APE1) (4128; Cell Signaling Technology), superoxide dismutase–1 (SOD1) (2770; Cell Signaling Technology), and 8-oxoguanine glycosylase 1 (OGG1) (sc33181; Santa Cruz Biotech), followed by HRP-conjugated secondary antibody.

    Techniques: Generated

    Probucol activated the Nrf2/ARE signaling pathways in astrocytes a . b . Immunofluorescence analysis was used to detect the staining intensity of GFAP and Nrf2. Scale bars are 50 μm. * P

    Journal: Oncotarget

    Article Title: Activation of the Nrf2/ARE signaling pathway by probucol contributes to inhibiting inflammation and neuronal apoptosis after spinal cord injury

    doi: 10.18632/oncotarget.19107

    Figure Lengend Snippet: Probucol activated the Nrf2/ARE signaling pathways in astrocytes a . b . Immunofluorescence analysis was used to detect the staining intensity of GFAP and Nrf2. Scale bars are 50 μm. * P

    Article Snippet: Subsequently, the separated proteins were transferred onto a polyvinylidene fluoride (PVDF) membrane and blocked with 5% skim milk powder in TBST (25 mM Tris-HCl, 0.15 M saline, and 1% Tween 20) at room temperature for 2 h. Membranes were then incubated overnight at 4°C with the following primary antibodies: NeuN (1:1,000, Cell Signaling Technology, Danvers, MA, USA), GFAP (1:1,000, Abcam, Cambridge, UK), Nrf2 (1:1,000, Cell Signaling Technology), HO-1 (1:1,000, Abcam), NQO1 (1:1,000, Abcam), pan-Akt (1:1,000, Abcam), p-Akt (1:1,000, Abcam), IL-1β (1:500, Abcam), IL-6 (1:2,000, Abcam), TNF-α (1:1,000, Abcam), Bcl-2 (1:1,000, Cell Signaling Technology), Bax (1:1,000, Cell Signaling Technology), cleaved caspase-3 (1:1,000, Cell Signaling Technology), cleaved PARP (1:1,000, Cell Signaling Technology), iNOS (1:1,000, Cell Signaling Technology), NF-κB (1:1,000, Cell Signaling Technology) and GAPDH (1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

    Techniques: Immunofluorescence, Staining

    Probucol activated the Akt/Nrf2/ARE signaling pathways after SCI in rat a . b . Representative western blots and quantification data of p-Akt, pan-Akt and GAPDH in each group rats on the seventh postoperative day; columns represent mean ± SD, * P

    Journal: Oncotarget

    Article Title: Activation of the Nrf2/ARE signaling pathway by probucol contributes to inhibiting inflammation and neuronal apoptosis after spinal cord injury

    doi: 10.18632/oncotarget.19107

    Figure Lengend Snippet: Probucol activated the Akt/Nrf2/ARE signaling pathways after SCI in rat a . b . Representative western blots and quantification data of p-Akt, pan-Akt and GAPDH in each group rats on the seventh postoperative day; columns represent mean ± SD, * P

    Article Snippet: Subsequently, the separated proteins were transferred onto a polyvinylidene fluoride (PVDF) membrane and blocked with 5% skim milk powder in TBST (25 mM Tris-HCl, 0.15 M saline, and 1% Tween 20) at room temperature for 2 h. Membranes were then incubated overnight at 4°C with the following primary antibodies: NeuN (1:1,000, Cell Signaling Technology, Danvers, MA, USA), GFAP (1:1,000, Abcam, Cambridge, UK), Nrf2 (1:1,000, Cell Signaling Technology), HO-1 (1:1,000, Abcam), NQO1 (1:1,000, Abcam), pan-Akt (1:1,000, Abcam), p-Akt (1:1,000, Abcam), IL-1β (1:500, Abcam), IL-6 (1:2,000, Abcam), TNF-α (1:1,000, Abcam), Bcl-2 (1:1,000, Cell Signaling Technology), Bax (1:1,000, Cell Signaling Technology), cleaved caspase-3 (1:1,000, Cell Signaling Technology), cleaved PARP (1:1,000, Cell Signaling Technology), iNOS (1:1,000, Cell Signaling Technology), NF-κB (1:1,000, Cell Signaling Technology) and GAPDH (1:1,000, Santa Cruz Biotechnology, Santa Cruz, CA, USA).

    Techniques: Western Blot

    Immunoblotting of NRF2, AMPK, HIF, evaluated on HS-68 cells exposed to 1 μmol/L BDE 209, 99, 47 and MIX for 12 and 20 days. Actin was used as internal control. The images are representative of at least three separate experiments. The relative protein quantification is represented in the graphic (* p

    Journal: International Journal of Environmental Research and Public Health

    Article Title: Sub-lethal Doses of Polybrominated Diphenyl Ethers, in Vitro, Promote Oxidative Stress and Modulate Molecular Markers Related to Cell Cycle, Antioxidant Balance and Cellular Energy Management

    doi: 10.3390/ijerph16040588

    Figure Lengend Snippet: Immunoblotting of NRF2, AMPK, HIF, evaluated on HS-68 cells exposed to 1 μmol/L BDE 209, 99, 47 and MIX for 12 and 20 days. Actin was used as internal control. The images are representative of at least three separate experiments. The relative protein quantification is represented in the graphic (* p

    Article Snippet: Filters were used for protein detection by primary antibodies (AbI) specifics for p53, retinoblastoma protein (pRB, IF8), Extracellular signal-regulated kinase 1 (ERK1), c-Fos, c-Jun, phospho-AMP-activated protein kinase (AMPK), hypoxia-inducible factor (HIF), poly (ADP-ribose) Polymerases (PARP) and Nuclear factor (erythroid-derived 2)-like 2 (NRF2) (Sigma-Aldrich Saint Louis, MO, USA).

    Techniques:

    The protective effects of the H 2 S donor on the expression of antioxidant-related proteins in the ageing kidney tissue. (a) The expression change of Nrf2 ( N = 11), HO-1 ( N = 10), SOD1 ( N = 11), and SOD2 ( N = 10) after 10 weeks of exogenous H 2 S donor. (b) Nrf2 was translocated from cytosol to nucleus, Lamin B was used as nuclear control, and GAPDH was used as cytosol control ( N = 6). Values are mean ± SE. P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Protective Effects of Hydrogen Sulfide in the Ageing Kidney

    doi: 10.1155/2016/7570489

    Figure Lengend Snippet: The protective effects of the H 2 S donor on the expression of antioxidant-related proteins in the ageing kidney tissue. (a) The expression change of Nrf2 ( N = 11), HO-1 ( N = 10), SOD1 ( N = 11), and SOD2 ( N = 10) after 10 weeks of exogenous H 2 S donor. (b) Nrf2 was translocated from cytosol to nucleus, Lamin B was used as nuclear control, and GAPDH was used as cytosol control ( N = 6). Values are mean ± SE. P

    Article Snippet: The proteins were resolved on a sodium dodecyl sulfate 10% polyacrylamide gel and transferred onto polyvinylidene fluoride membrane (Millipore, Bedford, MA, USA) and incubated with primary antibodies (1 : 1000 dilution) against Bcl-2, Bax, CSE, CBS, 3-MST, SIRT1 (Santa Cruz, CA, USA), Collagen I (Col I), Collagen III (Col III), Fibronectin (FN), SOD1, SOD2 (Abcam Company, USA), or Nrf2, HO-1 (Proteintech, China) at 4°C overnight.

    Techniques: Expressing

    Klotho improved nuclear translocation of Nrf2 in DOX-exposed myocardium and myocytes which was reversed by MAPKs inhibitors or siRNAs. ( A, B ) The left panels show the immunoblots of Nrf2 and Histone H3 in nuclear protein samples extracted from myocardium and myocytes. Columns on the right panels indicate the ratio of Nrf2/histone H3 in myocardium and cultured myocytes respectively. ( C ) The left panel demonstrates the captured fluorescent images of Nrf2, DAPI, and their merged images in cultured myocytes. Columns on the right part indicate the Nrf2 nuclear translocation rate in different groups. a Differences were significant when compared with control ( p

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: Klotho Improves Cardiac Function by Suppressing Reactive Oxygen Species (ROS) Mediated Apoptosis by Modulating Mapks/Nrf2 Signaling in Doxorubicin-Induced Cardiotoxicity

    doi: 10.12659/MSM.907449

    Figure Lengend Snippet: Klotho improved nuclear translocation of Nrf2 in DOX-exposed myocardium and myocytes which was reversed by MAPKs inhibitors or siRNAs. ( A, B ) The left panels show the immunoblots of Nrf2 and Histone H3 in nuclear protein samples extracted from myocardium and myocytes. Columns on the right panels indicate the ratio of Nrf2/histone H3 in myocardium and cultured myocytes respectively. ( C ) The left panel demonstrates the captured fluorescent images of Nrf2, DAPI, and their merged images in cultured myocytes. Columns on the right part indicate the Nrf2 nuclear translocation rate in different groups. a Differences were significant when compared with control ( p

    Article Snippet: The protein samples were transferred electrically to PVDF or NC membranes which were then incubated with primary antibodies against p38 MAPK (Cell Signaling Tech, 1: 1,000), phospho-p38 MAPK (p-p38 MAPK, Cell Signaling Tech, 1: 1,000), JNK (Abcam, 1: 500), phospho-JNK (p-JNK, Abcam, 1: 500), ERK1/2 (Cell Signaling Tech, 1: 1,000), phospho-ERK1/2 (p-ERK1/2, Cell Signaling Tech, 1: 1,000), Nrf2 (Invitrogen, 1: 500), HO1 (Santa Cruz, 1: 1,000), Prx1 (Santa Cruz, 1: 1,000), GAPDH (Santa Cruz, 1: 2,000) and Lamin A (Santa Cruz, 1: 1,000) at 4°C for 10 hours.

    Techniques: Translocation Assay, Western Blot, Cell Culture

    Schematic representation of the p62- Keap1- Nrf2 pathway in ovarian cancer cells with the treatment of VK3. Our study provides evidence that p62 promotes Nrf2 signaling through interacting with Keap1, which blocks VK3-induced apoptosis by inhibiting ROS production in SKOV3/DDP cells.

    Journal: Journal of Cancer

    Article Title: p62 Suppressed VK3-induced Oxidative Damage Through Keap1/Nrf2 Pathway In Human Ovarian Cancer Cells

    doi: 10.7150/jca.34423

    Figure Lengend Snippet: Schematic representation of the p62- Keap1- Nrf2 pathway in ovarian cancer cells with the treatment of VK3. Our study provides evidence that p62 promotes Nrf2 signaling through interacting with Keap1, which blocks VK3-induced apoptosis by inhibiting ROS production in SKOV3/DDP cells.

    Article Snippet: Antibodies used in this study were anti-p62 and caspase 3 (Abcam, Cambridge, MA, USA ), anti-β actin and anti-Keap1 (Proteintech, Chicago, IL, USA), anti-Nrf2 (ABclonal Biotechnology Co., Ltd.) and anti-Lamin A/C (Santa Cruz Biotechnology, Santa Cruz, CA, USA).

    Techniques:

    VK3 activates the Nrf2 pathway in SKOV3/DDP cells. (A) Both cells were treated as before. Nucleus extracts were subjected to immunoblot analysis with anti-Nrf2 and anti-LaminA/C. (B) Quantitation of nucleus Nrf2 protein level in (A). Data are presented as mean ± SD, n = 3. * P

    Journal: Journal of Cancer

    Article Title: p62 Suppressed VK3-induced Oxidative Damage Through Keap1/Nrf2 Pathway In Human Ovarian Cancer Cells

    doi: 10.7150/jca.34423

    Figure Lengend Snippet: VK3 activates the Nrf2 pathway in SKOV3/DDP cells. (A) Both cells were treated as before. Nucleus extracts were subjected to immunoblot analysis with anti-Nrf2 and anti-LaminA/C. (B) Quantitation of nucleus Nrf2 protein level in (A). Data are presented as mean ± SD, n = 3. * P

    Article Snippet: Antibodies used in this study were anti-p62 and caspase 3 (Abcam, Cambridge, MA, USA ), anti-β actin and anti-Keap1 (Proteintech, Chicago, IL, USA), anti-Nrf2 (ABclonal Biotechnology Co., Ltd.) and anti-Lamin A/C (Santa Cruz Biotechnology, Santa Cruz, CA, USA).

    Techniques: Quantitation Assay

    Knockdown of p62 inhibited the translocation of Nrf2 in SKOV3/DDP cells with VK3 treatment. (A) si-p62 or si-scrambled were transfected with SKOV3/DDP cells. After treated with 15 µM VK3 for 16h, cell lysates were subjected to immunoblot analysis (B) Cells were treated as (A), Immunofluorescence was performed with anti-Nrf2 antibodies and detected by fluorescence microscopy (scale bar, 25 µm).

    Journal: Journal of Cancer

    Article Title: p62 Suppressed VK3-induced Oxidative Damage Through Keap1/Nrf2 Pathway In Human Ovarian Cancer Cells

    doi: 10.7150/jca.34423

    Figure Lengend Snippet: Knockdown of p62 inhibited the translocation of Nrf2 in SKOV3/DDP cells with VK3 treatment. (A) si-p62 or si-scrambled were transfected with SKOV3/DDP cells. After treated with 15 µM VK3 for 16h, cell lysates were subjected to immunoblot analysis (B) Cells were treated as (A), Immunofluorescence was performed with anti-Nrf2 antibodies and detected by fluorescence microscopy (scale bar, 25 µm).

    Article Snippet: Antibodies used in this study were anti-p62 and caspase 3 (Abcam, Cambridge, MA, USA ), anti-β actin and anti-Keap1 (Proteintech, Chicago, IL, USA), anti-Nrf2 (ABclonal Biotechnology Co., Ltd.) and anti-Lamin A/C (Santa Cruz Biotechnology, Santa Cruz, CA, USA).

    Techniques: Translocation Assay, Transfection, Immunofluorescence, Fluorescence, Microscopy

    Effects of dietary resveratrol supplementation during gestation and lactation of sows on mRNA relative expression of Nrf2-regulated genes and pro-inflammatory cytokines in placenta. a , b mRNA relative expression of Nrf2-regulated genes; c mRNA relative expression of pro-inflammatory cytokines. Con, control treatment; Res, resveratrol treatment; SOD , superoxide dismutase; Gpx , glutathione peroxidase; CAT , catalase; NQO1 , NAD(P)H quinone dehydrogenase 1; HO1 , heme oxygenase 1; CYP1A1 , cytochrome P450 family 1 subfamily A member 1; TXNRD1 , thioredoxin reductase 1; GCLM , glutamate-cysteine ligase modifier; MGST1 , microsomal glutathione S-transferase 1; UGT1A1 , UDP glucuronosyl-transferase family 1 member A1; IL-1β , interleukin 1β; IL-6 , interleukin 6; IL-8 , interleukin 8; TNF-α , tumor necrosis factor α. All values are expressed as means ± SEM (n = 6). GAPDH was used as the internal control. * P

    Journal: Journal of Animal Science and Biotechnology

    Article Title: Dietary resveratrol improves antioxidant status of sows and piglets and regulates antioxidant gene expression in placenta by Keap1-Nrf2 pathway and Sirt1

    doi: 10.1186/s40104-018-0248-y

    Figure Lengend Snippet: Effects of dietary resveratrol supplementation during gestation and lactation of sows on mRNA relative expression of Nrf2-regulated genes and pro-inflammatory cytokines in placenta. a , b mRNA relative expression of Nrf2-regulated genes; c mRNA relative expression of pro-inflammatory cytokines. Con, control treatment; Res, resveratrol treatment; SOD , superoxide dismutase; Gpx , glutathione peroxidase; CAT , catalase; NQO1 , NAD(P)H quinone dehydrogenase 1; HO1 , heme oxygenase 1; CYP1A1 , cytochrome P450 family 1 subfamily A member 1; TXNRD1 , thioredoxin reductase 1; GCLM , glutamate-cysteine ligase modifier; MGST1 , microsomal glutathione S-transferase 1; UGT1A1 , UDP glucuronosyl-transferase family 1 member A1; IL-1β , interleukin 1β; IL-6 , interleukin 6; IL-8 , interleukin 8; TNF-α , tumor necrosis factor α. All values are expressed as means ± SEM (n = 6). GAPDH was used as the internal control. * P

    Article Snippet: After transfer, membranes were blocked for 1 h at room temperature in TBST (10 mmol/L Tris-HCl, pH 8.0, 150 mmol/L NaCl, 0.2% Tween-20) containing 5% non-fat milk powder and incubated overnight at 4 °C with 1:500 diluted primary antibodies, including GAPDH (Wanlei Biotechnology), Sirt1 (Wanlei Biotechnology), NFκB-p65 (Santa Cruz Biotechnology), phosphorylated NFκB-p65 (Ser536, Santa Cruz Biotechnology), Nrf2 (Abacm) and Keap1(Bioss) antibodies.

    Techniques: Expressing

    Effects of dietary resveratrol supplementation during gestation and lactation of sows on Sirt1, Nrf2-Keap1, NFκB-p65 and p-NFκB-p65 protein expression in placenta. Con, control treatment; Res, resveratrol treatment; a Sirt1 protein expression; b Keap1 and Nrf2 protein expression; c NFκB-p65 and p-NFκB-p65 (Ser536) protein expression; Con, control treatment; Res, resveratrol treatment; Nrf2, Nuclear factor E2-related factor 2; Keap1, Kelch-like ECH associated protein 1; NFκB, nuclear factor kappa B; p-NFκB, phosphorylated NFκB. All values are expressed as means ± SEM ( n = 6). GAPDH was used as the internal control. * P

    Journal: Journal of Animal Science and Biotechnology

    Article Title: Dietary resveratrol improves antioxidant status of sows and piglets and regulates antioxidant gene expression in placenta by Keap1-Nrf2 pathway and Sirt1

    doi: 10.1186/s40104-018-0248-y

    Figure Lengend Snippet: Effects of dietary resveratrol supplementation during gestation and lactation of sows on Sirt1, Nrf2-Keap1, NFκB-p65 and p-NFκB-p65 protein expression in placenta. Con, control treatment; Res, resveratrol treatment; a Sirt1 protein expression; b Keap1 and Nrf2 protein expression; c NFκB-p65 and p-NFκB-p65 (Ser536) protein expression; Con, control treatment; Res, resveratrol treatment; Nrf2, Nuclear factor E2-related factor 2; Keap1, Kelch-like ECH associated protein 1; NFκB, nuclear factor kappa B; p-NFκB, phosphorylated NFκB. All values are expressed as means ± SEM ( n = 6). GAPDH was used as the internal control. * P

    Article Snippet: After transfer, membranes were blocked for 1 h at room temperature in TBST (10 mmol/L Tris-HCl, pH 8.0, 150 mmol/L NaCl, 0.2% Tween-20) containing 5% non-fat milk powder and incubated overnight at 4 °C with 1:500 diluted primary antibodies, including GAPDH (Wanlei Biotechnology), Sirt1 (Wanlei Biotechnology), NFκB-p65 (Santa Cruz Biotechnology), phosphorylated NFκB-p65 (Ser536, Santa Cruz Biotechnology), Nrf2 (Abacm) and Keap1(Bioss) antibodies.

    Techniques: Expressing

    DHA enhances the nuclear translocation and DNA binding activity of Nrf2 in neurons. Primary neuronal cultures were treated with vehicle or DHA followed by OGD. A , B , Representative Western blots ( A ) and semiquantitative analyses ( B ) of Nrf2 levels, showing increased nuclear Nrf2 in DHA-treated groups (* p ≤ 0.05 vs control and vehicle-treated OGD groups at the same time points; # p ≤ 0.05 vs control groups, n = 3 per condition). C , D , Representative electrophoretic mobility shift assay ( C ) and semiquantitative analyses ( D ) of DNA binding of Nrf2 (* p ≤ 0.05 vs control and vehicle-treated OGD groups at the same time points; # p ≤ 0.05 vs control groups; and p ≤ 0.01 vs DHA-treated OGD at 24 h; n = 3 per condition). The specificity of DNA binding activity was determined by a competition assay, whereas threefold (C1) or 50-fold (C2) excess of cold probe was added as a specific competitor.

    Journal: The Journal of Neuroscience

    Article Title: Omega-3 Fatty Acids Protect the Brain against Ischemic Injury by Activating Nrf2 and Upregulating Heme Oxygenase 1

    doi: 10.1523/JNEUROSCI.4043-13.2014

    Figure Lengend Snippet: DHA enhances the nuclear translocation and DNA binding activity of Nrf2 in neurons. Primary neuronal cultures were treated with vehicle or DHA followed by OGD. A , B , Representative Western blots ( A ) and semiquantitative analyses ( B ) of Nrf2 levels, showing increased nuclear Nrf2 in DHA-treated groups (* p ≤ 0.05 vs control and vehicle-treated OGD groups at the same time points; # p ≤ 0.05 vs control groups, n = 3 per condition). C , D , Representative electrophoretic mobility shift assay ( C ) and semiquantitative analyses ( D ) of DNA binding of Nrf2 (* p ≤ 0.05 vs control and vehicle-treated OGD groups at the same time points; # p ≤ 0.05 vs control groups; and p ≤ 0.01 vs DHA-treated OGD at 24 h; n = 3 per condition). The specificity of DNA binding activity was determined by a competition assay, whereas threefold (C1) or 50-fold (C2) excess of cold probe was added as a specific competitor.

    Article Snippet: Equal amounts of protein samples were loaded and probed with antibodies recognizing HO-1 (1:3000; Enzo Life Science), Nrf2 (1:1000; Enzo Life Science), Keap1 (1:1000; Abcam), β-actin (1:8000; Millipore), or histone (1:1000; Cell Signaling Technology).

    Techniques: Translocation Assay, Binding Assay, Activity Assay, Western Blot, Electrophoretic Mobility Shift Assay, Competitive Binding Assay

    Potential mechanism of n -3 PUFA-mediated neuroprotection. 1 , Under physiological conditions, Nrf2 is physically “locked” with Keap1, which leads to proteasomal degradation of Nrf2 via the Cul3 (E3)-dependent pathway. 2 , Under oxidative stress conditions, such as brain ischemia and reperfusion, lipid oxidation produces α,β-unsaturated carbonyl electrophiles. n-3 PUFA-derived 4-HHE is more powerful than n-6 PUFA-derived 4-HNE in electrophilicity. 3 , Four-HHE covalently reacts with cysteine residues of Keap1, which causes conformational changes in Keap1, setting Nrf2 free. 4 , Nrf2 then accumulates, translocates into the nucleus, and binds the antioxidant response element (ARE) of phase 2 genes, leading to upregulation of HO-1 and other enzymes. 5 , HO-1 protects the brain against ischemic damage by breaking down heme into biliverdin (antioxidative) and carbon monoxide (CO; anti-inflammatory), reducing calcium overload and positive feedback on HO-1 expression. C, Cysteines; Cul3, cullin3; PLA2, phospholipase A2; Ub, ubiquitin.

    Journal: The Journal of Neuroscience

    Article Title: Omega-3 Fatty Acids Protect the Brain against Ischemic Injury by Activating Nrf2 and Upregulating Heme Oxygenase 1

    doi: 10.1523/JNEUROSCI.4043-13.2014

    Figure Lengend Snippet: Potential mechanism of n -3 PUFA-mediated neuroprotection. 1 , Under physiological conditions, Nrf2 is physically “locked” with Keap1, which leads to proteasomal degradation of Nrf2 via the Cul3 (E3)-dependent pathway. 2 , Under oxidative stress conditions, such as brain ischemia and reperfusion, lipid oxidation produces α,β-unsaturated carbonyl electrophiles. n-3 PUFA-derived 4-HHE is more powerful than n-6 PUFA-derived 4-HNE in electrophilicity. 3 , Four-HHE covalently reacts with cysteine residues of Keap1, which causes conformational changes in Keap1, setting Nrf2 free. 4 , Nrf2 then accumulates, translocates into the nucleus, and binds the antioxidant response element (ARE) of phase 2 genes, leading to upregulation of HO-1 and other enzymes. 5 , HO-1 protects the brain against ischemic damage by breaking down heme into biliverdin (antioxidative) and carbon monoxide (CO; anti-inflammatory), reducing calcium overload and positive feedback on HO-1 expression. C, Cysteines; Cul3, cullin3; PLA2, phospholipase A2; Ub, ubiquitin.

    Article Snippet: Equal amounts of protein samples were loaded and probed with antibodies recognizing HO-1 (1:3000; Enzo Life Science), Nrf2 (1:1000; Enzo Life Science), Keap1 (1:1000; Abcam), β-actin (1:8000; Millipore), or histone (1:1000; Cell Signaling Technology).

    Techniques: Derivative Assay, Expressing

    Ethanol induces Nrf2/ARE binding activity in PCN nuclear extracts. A, nuclear protein obtained from controls and PCNs exposed to ETOH (4 mg/ml) for different time points was used to detect ARE-specific oligonucleotide-protein complexes by EMSA. B, supershift

    Journal: Molecular Pharmacology

    Article Title: Overexpression of Nrf2 Protects Cerebral Cortical Neurons from Ethanol-Induced Apoptotic Death S⃞

    doi: 10.1124/mol.111.073262

    Figure Lengend Snippet: Ethanol induces Nrf2/ARE binding activity in PCN nuclear extracts. A, nuclear protein obtained from controls and PCNs exposed to ETOH (4 mg/ml) for different time points was used to detect ARE-specific oligonucleotide-protein complexes by EMSA. B, supershift

    Article Snippet: Subsequent supershift assays were performed using biotin-based EMSA with multiple newly available Nrf2 antibodies, out of which anti-Nrf2 (OriGene, Rockville, MD) produced a supershift (Supplemental Fig. 4).

    Techniques: Binding Assay, Activity Assay

    Adenovirus-mediated overexpression of Nrf2 contained ethanol-induced oxidative stress and neuronal death. A and B, PCNs (4DIV) in serum containing medium were infected with adenovirus encoding Nrf2 cDNA (A) or DN Nrf2 (B) at 200 MOI. Twenty-four and 48

    Journal: Molecular Pharmacology

    Article Title: Overexpression of Nrf2 Protects Cerebral Cortical Neurons from Ethanol-Induced Apoptotic Death S⃞

    doi: 10.1124/mol.111.073262

    Figure Lengend Snippet: Adenovirus-mediated overexpression of Nrf2 contained ethanol-induced oxidative stress and neuronal death. A and B, PCNs (4DIV) in serum containing medium were infected with adenovirus encoding Nrf2 cDNA (A) or DN Nrf2 (B) at 200 MOI. Twenty-four and 48

    Article Snippet: Subsequent supershift assays were performed using biotin-based EMSA with multiple newly available Nrf2 antibodies, out of which anti-Nrf2 (OriGene, Rockville, MD) produced a supershift (Supplemental Fig. 4).

    Techniques: Over Expression, Infection

    Ethanol induces Nrf2 expression in a time-dependent manner in primary cortical neurons. PCNs were treated with ETOH (4 mg/ml) for the times indicated. A, Nrf2 protein expression was determined by immunoblot analyses and equal loading demonstrated by anti-GAPDH.

    Journal: Molecular Pharmacology

    Article Title: Overexpression of Nrf2 Protects Cerebral Cortical Neurons from Ethanol-Induced Apoptotic Death S⃞

    doi: 10.1124/mol.111.073262

    Figure Lengend Snippet: Ethanol induces Nrf2 expression in a time-dependent manner in primary cortical neurons. PCNs were treated with ETOH (4 mg/ml) for the times indicated. A, Nrf2 protein expression was determined by immunoblot analyses and equal loading demonstrated by anti-GAPDH.

    Article Snippet: Subsequent supershift assays were performed using biotin-based EMSA with multiple newly available Nrf2 antibodies, out of which anti-Nrf2 (OriGene, Rockville, MD) produced a supershift (Supplemental Fig. 4).

    Techniques: Expressing

    siRNA-mediated Nrf2 knockdown exacerbates ethanol-induced oxidative stress and neuronal death. A, PCNs were transfected with either nontargeting scramble siRNA or a SMARTpool mix of four siNrf2s using siPORT amine. Twenty-four hours after transfection

    Journal: Molecular Pharmacology

    Article Title: Overexpression of Nrf2 Protects Cerebral Cortical Neurons from Ethanol-Induced Apoptotic Death S⃞

    doi: 10.1124/mol.111.073262

    Figure Lengend Snippet: siRNA-mediated Nrf2 knockdown exacerbates ethanol-induced oxidative stress and neuronal death. A, PCNs were transfected with either nontargeting scramble siRNA or a SMARTpool mix of four siNrf2s using siPORT amine. Twenty-four hours after transfection

    Article Snippet: Subsequent supershift assays were performed using biotin-based EMSA with multiple newly available Nrf2 antibodies, out of which anti-Nrf2 (OriGene, Rockville, MD) produced a supershift (Supplemental Fig. 4).

    Techniques: Transfection

    Prenatal ethanol exposure increases Nrf2 expression and activation in fetal brain cortices. Pregnant rats (Sprague-Dawley) at embryonic day 16 were administered ETOH (4 g/kg b.wt.) or isocaloric dextrose by gastric intubation at 12-h intervals for 2 days.

    Journal: Molecular Pharmacology

    Article Title: Overexpression of Nrf2 Protects Cerebral Cortical Neurons from Ethanol-Induced Apoptotic Death S⃞

    doi: 10.1124/mol.111.073262

    Figure Lengend Snippet: Prenatal ethanol exposure increases Nrf2 expression and activation in fetal brain cortices. Pregnant rats (Sprague-Dawley) at embryonic day 16 were administered ETOH (4 g/kg b.wt.) or isocaloric dextrose by gastric intubation at 12-h intervals for 2 days.

    Article Snippet: Subsequent supershift assays were performed using biotin-based EMSA with multiple newly available Nrf2 antibodies, out of which anti-Nrf2 (OriGene, Rockville, MD) produced a supershift (Supplemental Fig. 4).

    Techniques: Expressing, Activation Assay