mouse monoclonal anti-ho-1 antibody Search Results


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  • 93
    Thermo Fisher mouse anti hmox1
    Autophagy induction of autophagy-related genes by cigarette smoke extract in vitro paralleled by upregulation of OSGIN1. Primary human airway basal cells were treated with cigarette smoke extract (CSE) in vitro. Increasing concentrations of CSE (0 to 10%) were assessed (n = 3/group). (A) TaqMan PCR assessment of gene expression changes induced by CSE. Oxidative stress genes <t>HMOX1,</t> CYP1A1 and NQO1 were used as positive control. Shown are autophagy-related genes, MAP1LC3B, SQSTM1 and GABARAPL1 , paralleled by OSGIN1 expression. Values are mean ± standard deviation; *, P
    Mouse Anti Hmox1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Millipore mouse anti human ho 1 monoclonal antibody
    FerrylHb induces <t>HO-1</t> and ferritin in EC. Confluent HUVECs grown on 6-well plates were exposed to heme, Hb, metHb, and ferrylHb (100 μ mol/L heme or as indicated in complete medium containing 15% of FBS). After 4 hours of incubation total RNA was isolated and HO-1 mRNA level was measured by quantitative RT-PCR (panels (a) and (d)). For protein expression, HUVECs were solubilized after 8 hours of treatment. HO-1 and ferritin H and L expression was detected by Western blot ((b) and (e)) or with ELISA (c). Immunoblots were reprobed with GAPDH and are representative of three independent experiments. Results are shown as mean ± S.D. ( n = 3) from one representative experiment of three.
    Mouse Anti Human Ho 1 Monoclonal Antibody, supplied by Millipore, used in various techniques. Bioz Stars score: 89/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Santa Cruz Biotechnology mouse monoclonal anti ho 1 antibody
    Plate layout of one replicate of a <t>HO-1</t> measurement plate using the HTRF assay.
    Mouse Monoclonal Anti Ho 1 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse monoclonal anti ho 1 antibody/product/Santa Cruz Biotechnology
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    89
    Stressgen Biotechnologies mouse anti ho 1 monoclonal antibody
    Heme-oxygenase expression (HO-2 and <t>HO-1</t> expressed as %) in the cardiac left ventricle of male (the black square) and female (the grey square). Data are expressed as means ± S.E.M. of the results of a minimum of 10 rats per group. Statistical significance: * P
    Mouse Anti Ho 1 Monoclonal Antibody, supplied by Stressgen Biotechnologies, used in various techniques. Bioz Stars score: 89/100, based on 69 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam mouse anti hmox1 monoclonal antibody
    Changes in <t>HMOX1</t> expression following demethylation in H4 and H4-sw cells. H4 and H4-sw cells were treated with 10 μM 5-aza-2′-deoxycytidine for 3 days. After treatment, (A and C) HMOX1 mRNA expression was measured by qRT-PCR. (B and D) DNA methylation status at the −374 CpG site was measured using qMSP. Data are shown as mean ± SD (n = 3). Statistical analyses were performed using t -tests (** p
    Mouse Anti Hmox1 Monoclonal Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Becton Dickinson anti ho 1 mouse monoclonal antibody
    <t>HO-1</t> metabolites mediate resistance effects to chemotherapy . Cell growth was assessed 72 h after application of gemcitabine via MTT assay. (5A) Biliverdin administration biliverdin [5 and 50 μmol/l] resulted in increased proliferation of cancer cell line PANC-1 and S2-013, and therefore reduced the chemotherapeutic effect of gemcitabine (LD50 dose). (5B) Administration of ferrous histidinate revealed increased cell proliferation of PANC-1 and S2-013 cancer cells.
    Anti Ho 1 Mouse Monoclonal Antibody, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Cell Signaling Technology Inc anti ho 1 rabbit monoclonal antibody
    Effect of Nrf2 on GA-induced cytotoxicity. Schwann cells were treated with 500 μM GA for 24 h. (A) <t>HO-1</t> mRNA expression was analyzed by real-time RT-PCR. (B) HO-1 protein expression was analyzed under a laser scanning microscope. Scale bar, 20 μm. (C) Schwann cells were transfected with control siRNA (ctrl siRNA) or Nrf2 siRNA and were treated or not treated with GA. Data are means ± S.D. of three independent experiments. *Significant difference from the value of control ( p
    Anti Ho 1 Rabbit Monoclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 97/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Abcam phycoerytrin conjugated mouse monoclonal anti ho 1 antibody
    Effect of Nrf2 on GA-induced cytotoxicity. Schwann cells were treated with 500 μM GA for 24 h. (A) <t>HO-1</t> mRNA expression was analyzed by real-time RT-PCR. (B) HO-1 protein expression was analyzed under a laser scanning microscope. Scale bar, 20 μm. (C) Schwann cells were transfected with control siRNA (ctrl siRNA) or Nrf2 siRNA and were treated or not treated with GA. Data are means ± S.D. of three independent experiments. *Significant difference from the value of control ( p
    Phycoerytrin Conjugated Mouse Monoclonal Anti Ho 1 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 91/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Novus Biologicals monoclonal anti mouse ho 1 primary antibody
    ATF4 induces expression of <t>HO-1</t> in response to detachment-activated oxidative stress. ( A ) RT-PCR analysis of HO1 , GPX1 , and SOD2 in shNT and shATF4.HT1080 cells grown in suspension for 48 hours. Data are represented as mean fold change compared with attached cultures for 3 independent experiments ( n = 3, mean ± SD). * P
    Monoclonal Anti Mouse Ho 1 Primary Antibody, supplied by Novus Biologicals, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Santa Cruz Biotechnology mouse monoclonal anti ho 1 sc 390991 antibody
    ATF4 induces expression of <t>HO-1</t> in response to detachment-activated oxidative stress. ( A ) RT-PCR analysis of HO1 , GPX1 , and SOD2 in shNT and shATF4.HT1080 cells grown in suspension for 48 hours. Data are represented as mean fold change compared with attached cultures for 3 independent experiments ( n = 3, mean ± SD). * P
    Mouse Monoclonal Anti Ho 1 Sc 390991 Antibody, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Assay Designs Inc mouse monoclonal anti human ho 1 antibody
    Area under the curve (AUC) over 48 h showing changes from individual baseline levels (ΔAUC48) for haem oxygenase <t>(HO-1)</t> mRNA and protein levels. Box and whisker plots of the ΔAUC48 for the HO-1 mRNA (A) and of the ODs of the protein levels
    Mouse Monoclonal Anti Human Ho 1 Antibody, supplied by Assay Designs Inc, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Abcam unconjugated mouse rabbit anti ho 1 monoclonal antibody
    Area under the curve (AUC) over 48 h showing changes from individual baseline levels (ΔAUC48) for haem oxygenase <t>(HO-1)</t> mRNA and protein levels. Box and whisker plots of the ΔAUC48 for the HO-1 mRNA (A) and of the ODs of the protein levels
    Unconjugated Mouse Rabbit Anti Ho 1 Monoclonal Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Abcam mouse anti heme oxygenase 1 ho 1 monoclonal antibody
    Area under the curve (AUC) over 48 h showing changes from individual baseline levels (ΔAUC48) for haem oxygenase <t>(HO-1)</t> mRNA and protein levels. Box and whisker plots of the ΔAUC48 for the HO-1 mRNA (A) and of the ODs of the protein levels
    Mouse Anti Heme Oxygenase 1 Ho 1 Monoclonal Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 86/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Autophagy induction of autophagy-related genes by cigarette smoke extract in vitro paralleled by upregulation of OSGIN1. Primary human airway basal cells were treated with cigarette smoke extract (CSE) in vitro. Increasing concentrations of CSE (0 to 10%) were assessed (n = 3/group). (A) TaqMan PCR assessment of gene expression changes induced by CSE. Oxidative stress genes HMOX1, CYP1A1 and NQO1 were used as positive control. Shown are autophagy-related genes, MAP1LC3B, SQSTM1 and GABARAPL1 , paralleled by OSGIN1 expression. Values are mean ± standard deviation; *, P

    Journal: Autophagy

    Article Title: Role of OSGIN1 in mediating smoking-induced autophagy in the human airway epithelium

    doi: 10.1080/15548627.2017.1301327

    Figure Lengend Snippet: Autophagy induction of autophagy-related genes by cigarette smoke extract in vitro paralleled by upregulation of OSGIN1. Primary human airway basal cells were treated with cigarette smoke extract (CSE) in vitro. Increasing concentrations of CSE (0 to 10%) were assessed (n = 3/group). (A) TaqMan PCR assessment of gene expression changes induced by CSE. Oxidative stress genes HMOX1, CYP1A1 and NQO1 were used as positive control. Shown are autophagy-related genes, MAP1LC3B, SQSTM1 and GABARAPL1 , paralleled by OSGIN1 expression. Values are mean ± standard deviation; *, P

    Article Snippet: The following primary antibodies were used: rabbit anti-human MAP1LC3B (1:200 for immunofluorescence, 1:800 for immunohistochemistry; Cell Signaling Technology, 3868); mouse anti-human LAMP1 (1:100; R & D Systems, MAB4800); mouse Anti-SQSTM1 (1:2000; BD Biosciences, 610833); mouse anti-human OSGIN1 (1:100; Sigma-Aldrich, SAB1407392), mouse anti-CD63 (1:100; Santa Cruz Biotechnology, sc-5275), mouse anti-NQO1 (1:200; Abcam, ab28947), mouse anti-HMOX1 (1:100; Thermo Scientific, MA1–112).

    Techniques: In Vitro, Polymerase Chain Reaction, Expressing, Positive Control, Standard Deviation

    FerrylHb induces HO-1 and ferritin in EC. Confluent HUVECs grown on 6-well plates were exposed to heme, Hb, metHb, and ferrylHb (100 μ mol/L heme or as indicated in complete medium containing 15% of FBS). After 4 hours of incubation total RNA was isolated and HO-1 mRNA level was measured by quantitative RT-PCR (panels (a) and (d)). For protein expression, HUVECs were solubilized after 8 hours of treatment. HO-1 and ferritin H and L expression was detected by Western blot ((b) and (e)) or with ELISA (c). Immunoblots were reprobed with GAPDH and are representative of three independent experiments. Results are shown as mean ± S.D. ( n = 3) from one representative experiment of three.

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Atherogenesis May Involve the Prooxidant and Proinflammatory Effects of Ferryl Hemoglobin

    doi: 10.1155/2013/676425

    Figure Lengend Snippet: FerrylHb induces HO-1 and ferritin in EC. Confluent HUVECs grown on 6-well plates were exposed to heme, Hb, metHb, and ferrylHb (100 μ mol/L heme or as indicated in complete medium containing 15% of FBS). After 4 hours of incubation total RNA was isolated and HO-1 mRNA level was measured by quantitative RT-PCR (panels (a) and (d)). For protein expression, HUVECs were solubilized after 8 hours of treatment. HO-1 and ferritin H and L expression was detected by Western blot ((b) and (e)) or with ELISA (c). Immunoblots were reprobed with GAPDH and are representative of three independent experiments. Results are shown as mean ± S.D. ( n = 3) from one representative experiment of three.

    Article Snippet: After electrophoresis, proteins were transferred to a nitrocellulose membrane (Amersham Biosciences Corp., Piscataway, NJ, USA) and HO-1 was identified using a mouse anti-human HO-1 monoclonal antibody (Cat no. 374087, Calbiochem, Merck KGaA, Darmstadt, Germany) at a dilution of 1 : 2500.

    Techniques: Incubation, Isolation, Quantitative RT-PCR, Expressing, Western Blot, Enzyme-linked Immunosorbent Assay

    Plate layout of one replicate of a HO-1 measurement plate using the HTRF assay.

    Journal: Current protocols in toxicology / editorial board, Mahin D. Maines (editor-in-chief) ... [et al.]

    Article Title: A high-throughput screening assay to identify kidney toxic compounds

    doi: 10.1002/cptx.12

    Figure Lengend Snippet: Plate layout of one replicate of a HO-1 measurement plate using the HTRF assay.

    Article Snippet: Paraformaldehyde (PFA) (37%) (Sigma) Methanol (Sigma) EL 406 washer dispenser & Biostack 3 microplate stacker (Bio-Tek) Straight 24-place stainless steel Manifold for Microtest Plates for 384-well plates (Drummond) Odyssey blocking buffer (LiCor) Mouse monoclonal anti-HO-1 antibody (Santa Cruz) Goat anti-mouse antibody (Alexa 555) (Life Technologies) Mouse IgG Isotype control (Thermo Fisher Scientific) Microseal Foil for IF (BioRad) Operetta high-content microscope with plate handler (Perkin Elmer)

    Techniques: HTRF Assay

    Representative results of HO-1 measurements using the HTRF assay. The positive control (50 μM CdCl2) and three kidney toxic compounds (cisplatin, citrinin, and arsenic trioxide) show dose-depend, significant increase in HO-1 fold change compared

    Journal: Current protocols in toxicology / editorial board, Mahin D. Maines (editor-in-chief) ... [et al.]

    Article Title: A high-throughput screening assay to identify kidney toxic compounds

    doi: 10.1002/cptx.12

    Figure Lengend Snippet: Representative results of HO-1 measurements using the HTRF assay. The positive control (50 μM CdCl2) and three kidney toxic compounds (cisplatin, citrinin, and arsenic trioxide) show dose-depend, significant increase in HO-1 fold change compared

    Article Snippet: Paraformaldehyde (PFA) (37%) (Sigma) Methanol (Sigma) EL 406 washer dispenser & Biostack 3 microplate stacker (Bio-Tek) Straight 24-place stainless steel Manifold for Microtest Plates for 384-well plates (Drummond) Odyssey blocking buffer (LiCor) Mouse monoclonal anti-HO-1 antibody (Santa Cruz) Goat anti-mouse antibody (Alexa 555) (Life Technologies) Mouse IgG Isotype control (Thermo Fisher Scientific) Microseal Foil for IF (BioRad) Operetta high-content microscope with plate handler (Perkin Elmer)

    Techniques: HTRF Assay, Positive Control

    Increased sensitivity in detection of CdCl 2 -induced toxicity by the HO-1 measurement compared with the standard ATP-based cell viability assay. Red line indicates lowest concentration of CdCl 2 with significant increase/ decrease of the readout compared

    Journal: Current protocols in toxicology / editorial board, Mahin D. Maines (editor-in-chief) ... [et al.]

    Article Title: A high-throughput screening assay to identify kidney toxic compounds

    doi: 10.1002/cptx.12

    Figure Lengend Snippet: Increased sensitivity in detection of CdCl 2 -induced toxicity by the HO-1 measurement compared with the standard ATP-based cell viability assay. Red line indicates lowest concentration of CdCl 2 with significant increase/ decrease of the readout compared

    Article Snippet: Paraformaldehyde (PFA) (37%) (Sigma) Methanol (Sigma) EL 406 washer dispenser & Biostack 3 microplate stacker (Bio-Tek) Straight 24-place stainless steel Manifold for Microtest Plates for 384-well plates (Drummond) Odyssey blocking buffer (LiCor) Mouse monoclonal anti-HO-1 antibody (Santa Cruz) Goat anti-mouse antibody (Alexa 555) (Life Technologies) Mouse IgG Isotype control (Thermo Fisher Scientific) Microseal Foil for IF (BioRad) Operetta high-content microscope with plate handler (Perkin Elmer)

    Techniques: Viability Assay, Concentration Assay

    Heme-oxygenase expression (HO-2 and HO-1 expressed as %) in the cardiac left ventricle of male (the black square) and female (the grey square). Data are expressed as means ± S.E.M. of the results of a minimum of 10 rats per group. Statistical significance: * P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Sexual Dimorphism of Cardiovascular Ischemia Susceptibility Is Mediated by Heme Oxygenase

    doi: 10.1155/2013/521563

    Figure Lengend Snippet: Heme-oxygenase expression (HO-2 and HO-1 expressed as %) in the cardiac left ventricle of male (the black square) and female (the grey square). Data are expressed as means ± S.E.M. of the results of a minimum of 10 rats per group. Statistical significance: * P

    Article Snippet: Two hours after blocking with PBS (pH 7.4), 0.25% tween 20, and 5.0% fat-free dried milk, the membrane was probed with mouse anti-HO-1 monoclonal antibody (1/10.000; 2 h) (StressGen Biotechnologies Corp., Victoria, Canada) or anti-HO-2 monoclonal antibody (1/1000; 2 h) (StressGen Biotechnologies Corp., Victoria, Canada) at room temperature, washed 3 times with PBS-tween 20 and then incubated with horseradish peroxidase-conjugated bovine antimouse antibody (1/2000; 1 h; Santa Cruz Biotechnology Inc., Santa Cruz, Ca, USA) for 1 h at room temperature.

    Techniques: Expressing

    Heme-oxygenase expression (HO2 and HO-1 expressed as %) in the aortic of male (the black square) and female (the grey square). Data are expressed as means ± S.E.M. of the results of a minimum of 10 rats per group. Statistical significance: * P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Sexual Dimorphism of Cardiovascular Ischemia Susceptibility Is Mediated by Heme Oxygenase

    doi: 10.1155/2013/521563

    Figure Lengend Snippet: Heme-oxygenase expression (HO2 and HO-1 expressed as %) in the aortic of male (the black square) and female (the grey square). Data are expressed as means ± S.E.M. of the results of a minimum of 10 rats per group. Statistical significance: * P

    Article Snippet: Two hours after blocking with PBS (pH 7.4), 0.25% tween 20, and 5.0% fat-free dried milk, the membrane was probed with mouse anti-HO-1 monoclonal antibody (1/10.000; 2 h) (StressGen Biotechnologies Corp., Victoria, Canada) or anti-HO-2 monoclonal antibody (1/1000; 2 h) (StressGen Biotechnologies Corp., Victoria, Canada) at room temperature, washed 3 times with PBS-tween 20 and then incubated with horseradish peroxidase-conjugated bovine antimouse antibody (1/2000; 1 h; Santa Cruz Biotechnology Inc., Santa Cruz, Ca, USA) for 1 h at room temperature.

    Techniques: Expressing

    Experimental design: in vitro , ex vivo , in vivo measurements from heart aorta in intact female (in the proestrus phase) and male Wistar rats HO-1: heme oxygenase 1; HO-2: heme oxygenase 2; AVP: Arginine vasopressin.

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Sexual Dimorphism of Cardiovascular Ischemia Susceptibility Is Mediated by Heme Oxygenase

    doi: 10.1155/2013/521563

    Figure Lengend Snippet: Experimental design: in vitro , ex vivo , in vivo measurements from heart aorta in intact female (in the proestrus phase) and male Wistar rats HO-1: heme oxygenase 1; HO-2: heme oxygenase 2; AVP: Arginine vasopressin.

    Article Snippet: Two hours after blocking with PBS (pH 7.4), 0.25% tween 20, and 5.0% fat-free dried milk, the membrane was probed with mouse anti-HO-1 monoclonal antibody (1/10.000; 2 h) (StressGen Biotechnologies Corp., Victoria, Canada) or anti-HO-2 monoclonal antibody (1/1000; 2 h) (StressGen Biotechnologies Corp., Victoria, Canada) at room temperature, washed 3 times with PBS-tween 20 and then incubated with horseradish peroxidase-conjugated bovine antimouse antibody (1/2000; 1 h; Santa Cruz Biotechnology Inc., Santa Cruz, Ca, USA) for 1 h at room temperature.

    Techniques: In Vitro, Ex Vivo, In Vivo

    DEP and ox-PAPC co-regulate genes in a synergistic/additive fashion. (a) Synergistic index (SI). Synergy was defined as the presence of a co-regulatory effect by both DEP and ox-PAPC that was greater than the effects induced by either compound alone and greater than the sum of those individual effects. The following criteria for a synergistic effect were as follows: SI (ΔAB/(ΔA+ΔB) > 1; AB (mean) > A (mean), p ≤ 0.05; AB (mean) > B (mean), p ≤ 0.05, where ΔA is the difference in mean expression level between the DEP and the control samples, ΔB is the difference in mean expression level between the ox-PAPC and the control samples, and ΔAB is the difference in mean expression level between the DEP + ox-PAPC and the control samples. (b) mRNA expression levels of representative genes. Each graph displays the relative mRNA expression levels normalized by β 2 -microglobulin mRNA levels and expressed as fold control (FOC) for microarray (white bars, left-hand y -axis) and qPCR (black bars, right-hand y -axis) assessment of representative genes ( HO-1 , IL-8 , ATF4 , CXCL1 , XBP1 , IL-11 ). For ease of comparison, the qPCR scale was divided by factors of 3.5 ( HO-1 ) and 3 ( IL-8 ), respectively. In similar fashion, the microarray scale was divided by a factor of 4 ( IL-11 ) to make the comparison easier. The asterisk indicates combinations of DEP + ox-PAPC that exhibited synergistic effects. The high consistence of microarray and qPCR analysis, conducted on triplicate samples from independent experiments, implies both technical and biological validation. Statistical analysis was performed by one-way ANOVA, Fisher PLSD.

    Journal: Genome Biology

    Article Title: Air-pollutant chemicals and oxidized lipids exhibit genome-wide synergistic effects on endothelial cells

    doi: 10.1186/gb-2007-8-7-r149

    Figure Lengend Snippet: DEP and ox-PAPC co-regulate genes in a synergistic/additive fashion. (a) Synergistic index (SI). Synergy was defined as the presence of a co-regulatory effect by both DEP and ox-PAPC that was greater than the effects induced by either compound alone and greater than the sum of those individual effects. The following criteria for a synergistic effect were as follows: SI (ΔAB/(ΔA+ΔB) > 1; AB (mean) > A (mean), p ≤ 0.05; AB (mean) > B (mean), p ≤ 0.05, where ΔA is the difference in mean expression level between the DEP and the control samples, ΔB is the difference in mean expression level between the ox-PAPC and the control samples, and ΔAB is the difference in mean expression level between the DEP + ox-PAPC and the control samples. (b) mRNA expression levels of representative genes. Each graph displays the relative mRNA expression levels normalized by β 2 -microglobulin mRNA levels and expressed as fold control (FOC) for microarray (white bars, left-hand y -axis) and qPCR (black bars, right-hand y -axis) assessment of representative genes ( HO-1 , IL-8 , ATF4 , CXCL1 , XBP1 , IL-11 ). For ease of comparison, the qPCR scale was divided by factors of 3.5 ( HO-1 ) and 3 ( IL-8 ), respectively. In similar fashion, the microarray scale was divided by a factor of 4 ( IL-11 ) to make the comparison easier. The asterisk indicates combinations of DEP + ox-PAPC that exhibited synergistic effects. The high consistence of microarray and qPCR analysis, conducted on triplicate samples from independent experiments, implies both technical and biological validation. Statistical analysis was performed by one-way ANOVA, Fisher PLSD.

    Article Snippet: Mouse monoclonal anti-HO-1 antibody (StressGen Biotech, Victoria, Canada) and mouse monoclonal anti-β-actin antibody (Abcam, Cambridge, MA) were used as primary antibodies at 1:1,000 dilution overnight, respectively.

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

    Ambient ultrafine PM chemicals enhance in vivo expression of genes related to vascular inflammation. (a) Experimental protocol. Two-month-old male apoE null mice fed a high-fat diet were exposed for a total of 120 h (5 h/day, 3 days/week for 8 weeks) to concentrated ultrafine particles (UFP), concentrated PM 2.5 (FP), filtered air (FA) or not exposed (NE). (b) Hepatic gene expression levels. Gene expression was determined by qPCR of mRNA prepared from liver homogenates. UFP-exposed mice exhibited marked upregulation of HO-1 (left), XBP1 (center) and ATF4 (right). Values were normalized by β-actin mRNA levels and expressed as fold control (FOC). Five samples per group were assayed in duplicates. Statistical analysis was performed by one-way ANOVA, Fisher PLSD.

    Journal: Genome Biology

    Article Title: Air-pollutant chemicals and oxidized lipids exhibit genome-wide synergistic effects on endothelial cells

    doi: 10.1186/gb-2007-8-7-r149

    Figure Lengend Snippet: Ambient ultrafine PM chemicals enhance in vivo expression of genes related to vascular inflammation. (a) Experimental protocol. Two-month-old male apoE null mice fed a high-fat diet were exposed for a total of 120 h (5 h/day, 3 days/week for 8 weeks) to concentrated ultrafine particles (UFP), concentrated PM 2.5 (FP), filtered air (FA) or not exposed (NE). (b) Hepatic gene expression levels. Gene expression was determined by qPCR of mRNA prepared from liver homogenates. UFP-exposed mice exhibited marked upregulation of HO-1 (left), XBP1 (center) and ATF4 (right). Values were normalized by β-actin mRNA levels and expressed as fold control (FOC). Five samples per group were assayed in duplicates. Statistical analysis was performed by one-way ANOVA, Fisher PLSD.

    Article Snippet: Mouse monoclonal anti-HO-1 antibody (StressGen Biotech, Victoria, Canada) and mouse monoclonal anti-β-actin antibody (Abcam, Cambridge, MA) were used as primary antibodies at 1:1,000 dilution overnight, respectively.

    Techniques: In Vivo, Expressing, Mouse Assay, Real-time Polymerase Chain Reaction

    DEP and ox-PAPC induce HO-1 expression in HMEC. (a) Western blot. HMEC were treated with DEP, ox-PAPC or a combination of both at various concentrations. Mouse monoclonal anti-HO-1 and anti-β-actin antibodies were used to detect the relevant proteins as described in Materials and methods. (b) Densitometric analysis. The expression level of HO-1 protein in optical density (OD) units is shown. Similar levels of β-actin are shown in (a). Results are typical of one representative experiment ( n = 4).

    Journal: Genome Biology

    Article Title: Air-pollutant chemicals and oxidized lipids exhibit genome-wide synergistic effects on endothelial cells

    doi: 10.1186/gb-2007-8-7-r149

    Figure Lengend Snippet: DEP and ox-PAPC induce HO-1 expression in HMEC. (a) Western blot. HMEC were treated with DEP, ox-PAPC or a combination of both at various concentrations. Mouse monoclonal anti-HO-1 and anti-β-actin antibodies were used to detect the relevant proteins as described in Materials and methods. (b) Densitometric analysis. The expression level of HO-1 protein in optical density (OD) units is shown. Similar levels of β-actin are shown in (a). Results are typical of one representative experiment ( n = 4).

    Article Snippet: Mouse monoclonal anti-HO-1 antibody (StressGen Biotech, Victoria, Canada) and mouse monoclonal anti-β-actin antibody (Abcam, Cambridge, MA) were used as primary antibodies at 1:1,000 dilution overnight, respectively.

    Techniques: Expressing, Western Blot

    Changes in HMOX1 expression following demethylation in H4 and H4-sw cells. H4 and H4-sw cells were treated with 10 μM 5-aza-2′-deoxycytidine for 3 days. After treatment, (A and C) HMOX1 mRNA expression was measured by qRT-PCR. (B and D) DNA methylation status at the −374 CpG site was measured using qMSP. Data are shown as mean ± SD (n = 3). Statistical analyses were performed using t -tests (** p

    Journal: PLoS ONE

    Article Title: Amyloid Beta-Mediated Hypomethylation of Heme Oxygenase 1 Correlates with Cognitive Impairment in Alzheimer’s Disease

    doi: 10.1371/journal.pone.0153156

    Figure Lengend Snippet: Changes in HMOX1 expression following demethylation in H4 and H4-sw cells. H4 and H4-sw cells were treated with 10 μM 5-aza-2′-deoxycytidine for 3 days. After treatment, (A and C) HMOX1 mRNA expression was measured by qRT-PCR. (B and D) DNA methylation status at the −374 CpG site was measured using qMSP. Data are shown as mean ± SD (n = 3). Statistical analyses were performed using t -tests (** p

    Article Snippet: Membranes were blocked in 5% skim milk in Tris-buffered saline with 0.1% Tween 20 and incubated overnight at 4°C with the following primary antibodies: mouse anti-HMOX1 polyclonal antibody (1:1,000, Sigma-Aldrich, SAB14059), mouse anti-HMOX1 monoclonal antibody (1:250, abcam, ab13248) or mouse anti-β-actin monoclonal antibody (1:2,000, Santa Cruz Biotechnology, sc-47778).

    Techniques: Expressing, Quantitative RT-PCR, DNA Methylation Assay

    Evaluation of DNA methylation status at the –374 CpG site on the HMOX1 gene as a biomarker for AD. DNA methylation at the –374 CpG site in blood samples from AD patients, MCI patients and control individuals was analyzed using qMSP. (A) Data are expressed as scattered plots with lines at mean value. Statistical analyses were performed using Kruskal-Wallis one-way analysis of variance and Dunn’s multiple comparison post-tests for comparing the significance between AD patients and control individuals (*) or MCI patients ( # ) (*, # p

    Journal: PLoS ONE

    Article Title: Amyloid Beta-Mediated Hypomethylation of Heme Oxygenase 1 Correlates with Cognitive Impairment in Alzheimer’s Disease

    doi: 10.1371/journal.pone.0153156

    Figure Lengend Snippet: Evaluation of DNA methylation status at the –374 CpG site on the HMOX1 gene as a biomarker for AD. DNA methylation at the –374 CpG site in blood samples from AD patients, MCI patients and control individuals was analyzed using qMSP. (A) Data are expressed as scattered plots with lines at mean value. Statistical analyses were performed using Kruskal-Wallis one-way analysis of variance and Dunn’s multiple comparison post-tests for comparing the significance between AD patients and control individuals (*) or MCI patients ( # ) (*, # p

    Article Snippet: Membranes were blocked in 5% skim milk in Tris-buffered saline with 0.1% Tween 20 and incubated overnight at 4°C with the following primary antibodies: mouse anti-HMOX1 polyclonal antibody (1:1,000, Sigma-Aldrich, SAB14059), mouse anti-HMOX1 monoclonal antibody (1:250, abcam, ab13248) or mouse anti-β-actin monoclonal antibody (1:2,000, Santa Cruz Biotechnology, sc-47778).

    Techniques: DNA Methylation Assay, Biomarker Assay

    DNA methylation is altered in H4 cells treated with Aβ 1–42 or Aβ 1–40. (A) H4 cells were treated with 1 μM Aβ 1–42 or Aβ 1–40 for 5 days, and DNA methylation at the –374 CpG site was analyzed using qMSP. (B) After treatment with Aβ 1–42 or Aβ 1–40, HMOX1 expression was assessed using qPCR. (C) Protein levels of HMOX1 were determined by western blot analyses. Values shown are relative to untreated controls. Data are shown as mean ± SD (n = 3). Statistical analyses were performed using t -tests (** p

    Journal: PLoS ONE

    Article Title: Amyloid Beta-Mediated Hypomethylation of Heme Oxygenase 1 Correlates with Cognitive Impairment in Alzheimer’s Disease

    doi: 10.1371/journal.pone.0153156

    Figure Lengend Snippet: DNA methylation is altered in H4 cells treated with Aβ 1–42 or Aβ 1–40. (A) H4 cells were treated with 1 μM Aβ 1–42 or Aβ 1–40 for 5 days, and DNA methylation at the –374 CpG site was analyzed using qMSP. (B) After treatment with Aβ 1–42 or Aβ 1–40, HMOX1 expression was assessed using qPCR. (C) Protein levels of HMOX1 were determined by western blot analyses. Values shown are relative to untreated controls. Data are shown as mean ± SD (n = 3). Statistical analyses were performed using t -tests (** p

    Article Snippet: Membranes were blocked in 5% skim milk in Tris-buffered saline with 0.1% Tween 20 and incubated overnight at 4°C with the following primary antibodies: mouse anti-HMOX1 polyclonal antibody (1:1,000, Sigma-Aldrich, SAB14059), mouse anti-HMOX1 monoclonal antibody (1:250, abcam, ab13248) or mouse anti-β-actin monoclonal antibody (1:2,000, Santa Cruz Biotechnology, sc-47778).

    Techniques: DNA Methylation Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot

    HMOX1 expression is up-regulated in an APP-mutant cell line and transgenic mice. (A) HMOX1 mRNA expression in H4 and H4-sw cells was measured by transcriptome sequencing analyses and qRT-PCR. (B) Expression of HMOX1 protein in H4 and H4-sw cells was assessed using western blot analyses. Level of HMOX1 protein in H4-sw cells was expressed relative to that in H4 cells. (C) HMOX1 mRNA expression in the brain of 12-month-old wild-type and PSEN1-APP transgenic mice (B6C3-Tg(APP695)85Dbo Tg(PSEN1)85Dbo) was measured by qRT-PCR. (D) HMOX1 protein expression in the brain of 12-month-old wild-type and PSEN1-APP transgenic mice assessed using western blot analyses. Data are shown as mean ± SD. Statistical analyses were performed using t -tests (* p

    Journal: PLoS ONE

    Article Title: Amyloid Beta-Mediated Hypomethylation of Heme Oxygenase 1 Correlates with Cognitive Impairment in Alzheimer’s Disease

    doi: 10.1371/journal.pone.0153156

    Figure Lengend Snippet: HMOX1 expression is up-regulated in an APP-mutant cell line and transgenic mice. (A) HMOX1 mRNA expression in H4 and H4-sw cells was measured by transcriptome sequencing analyses and qRT-PCR. (B) Expression of HMOX1 protein in H4 and H4-sw cells was assessed using western blot analyses. Level of HMOX1 protein in H4-sw cells was expressed relative to that in H4 cells. (C) HMOX1 mRNA expression in the brain of 12-month-old wild-type and PSEN1-APP transgenic mice (B6C3-Tg(APP695)85Dbo Tg(PSEN1)85Dbo) was measured by qRT-PCR. (D) HMOX1 protein expression in the brain of 12-month-old wild-type and PSEN1-APP transgenic mice assessed using western blot analyses. Data are shown as mean ± SD. Statistical analyses were performed using t -tests (* p

    Article Snippet: Membranes were blocked in 5% skim milk in Tris-buffered saline with 0.1% Tween 20 and incubated overnight at 4°C with the following primary antibodies: mouse anti-HMOX1 polyclonal antibody (1:1,000, Sigma-Aldrich, SAB14059), mouse anti-HMOX1 monoclonal antibody (1:250, abcam, ab13248) or mouse anti-β-actin monoclonal antibody (1:2,000, Santa Cruz Biotechnology, sc-47778).

    Techniques: Expressing, Mutagenesis, Transgenic Assay, Mouse Assay, Sequencing, Quantitative RT-PCR, Western Blot

    Alterations of DNA methylation induced by Aβ are cell-types specific. Human leukemia cells (Jurkat, U937, HL60 and THP-1) were treated with 1 μM Aβ 1–42 for 6 days. After treatment with Aβ 1–42 (A) HMOX1 mRNA expression was assessed using qPCR, (B) DNA methylation of HMOX1 at the –374 CpG site was analyzed using qMSP. Values shown are relative to untreated controls. Data are shown as mean ± SD (n = 3). Statistical analyses were performed using t -tests (** p

    Journal: PLoS ONE

    Article Title: Amyloid Beta-Mediated Hypomethylation of Heme Oxygenase 1 Correlates with Cognitive Impairment in Alzheimer’s Disease

    doi: 10.1371/journal.pone.0153156

    Figure Lengend Snippet: Alterations of DNA methylation induced by Aβ are cell-types specific. Human leukemia cells (Jurkat, U937, HL60 and THP-1) were treated with 1 μM Aβ 1–42 for 6 days. After treatment with Aβ 1–42 (A) HMOX1 mRNA expression was assessed using qPCR, (B) DNA methylation of HMOX1 at the –374 CpG site was analyzed using qMSP. Values shown are relative to untreated controls. Data are shown as mean ± SD (n = 3). Statistical analyses were performed using t -tests (** p

    Article Snippet: Membranes were blocked in 5% skim milk in Tris-buffered saline with 0.1% Tween 20 and incubated overnight at 4°C with the following primary antibodies: mouse anti-HMOX1 polyclonal antibody (1:1,000, Sigma-Aldrich, SAB14059), mouse anti-HMOX1 monoclonal antibody (1:250, abcam, ab13248) or mouse anti-β-actin monoclonal antibody (1:2,000, Santa Cruz Biotechnology, sc-47778).

    Techniques: DNA Methylation Assay, Expressing, Real-time Polymerase Chain Reaction

    Hypomethylation of specific CpG sites within the HMOX1 promoter in H4-sw cells. DNA methylation status at the −374 CpG site was analyzed using (A) the Illumina HumanMethylation 27 BeadChip, (A) qMSP and (B) the 454 GS-FLX system. (C) The HMOX1 promoter region is located at position 35,380,082–35,381,037 in the human GRCh38/hg38 assembly and contains 17 CpG residues within chromosome 22. The 17 CpGs are located at positions −959, −919, −876, −791, −743, −706, −628, −602, −374, −341, −97, −92, −83, −81, −71, −59, and −55 from the transcription start site. Each circle represents CpG dinucleotides. The methylation status of each CpG site is illustrated by black (methylated) and white (unmethylated) circles, and the total percentage of methylation at each site is indicated by a pie graph on the top line. The black area of the pie graph indicates the methylated CpG percentage, whereas the white area indicates the unmethylated CpG percentage. Triangles above the circles or the bar in C indicate the specific CpG site that was used for qMSP and methylation microarray. Statistical analyses were performed using t -tests (*** p

    Journal: PLoS ONE

    Article Title: Amyloid Beta-Mediated Hypomethylation of Heme Oxygenase 1 Correlates with Cognitive Impairment in Alzheimer’s Disease

    doi: 10.1371/journal.pone.0153156

    Figure Lengend Snippet: Hypomethylation of specific CpG sites within the HMOX1 promoter in H4-sw cells. DNA methylation status at the −374 CpG site was analyzed using (A) the Illumina HumanMethylation 27 BeadChip, (A) qMSP and (B) the 454 GS-FLX system. (C) The HMOX1 promoter region is located at position 35,380,082–35,381,037 in the human GRCh38/hg38 assembly and contains 17 CpG residues within chromosome 22. The 17 CpGs are located at positions −959, −919, −876, −791, −743, −706, −628, −602, −374, −341, −97, −92, −83, −81, −71, −59, and −55 from the transcription start site. Each circle represents CpG dinucleotides. The methylation status of each CpG site is illustrated by black (methylated) and white (unmethylated) circles, and the total percentage of methylation at each site is indicated by a pie graph on the top line. The black area of the pie graph indicates the methylated CpG percentage, whereas the white area indicates the unmethylated CpG percentage. Triangles above the circles or the bar in C indicate the specific CpG site that was used for qMSP and methylation microarray. Statistical analyses were performed using t -tests (*** p

    Article Snippet: Membranes were blocked in 5% skim milk in Tris-buffered saline with 0.1% Tween 20 and incubated overnight at 4°C with the following primary antibodies: mouse anti-HMOX1 polyclonal antibody (1:1,000, Sigma-Aldrich, SAB14059), mouse anti-HMOX1 monoclonal antibody (1:250, abcam, ab13248) or mouse anti-β-actin monoclonal antibody (1:2,000, Santa Cruz Biotechnology, sc-47778).

    Techniques: DNA Methylation Assay, Methylation, Microarray

    PHB Tg mice are less susceptible to 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis independently of Nrf2. A : percent change in body weight. B : neutrophil infiltration into the colon, quantified by MPO activity. C : qRT-PCR analysis of HO-1, NQO-1, and Nrf2 in total RNA isolated from colon. Values are means ± SE ( n = 8 per group). * P

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Nrf2 is not required for epithelial prohibitin-dependent attenuation of experimental colitis

    doi: 10.1152/ajpgi.00327.2012

    Figure Lengend Snippet: PHB Tg mice are less susceptible to 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis independently of Nrf2. A : percent change in body weight. B : neutrophil infiltration into the colon, quantified by MPO activity. C : qRT-PCR analysis of HO-1, NQO-1, and Nrf2 in total RNA isolated from colon. Values are means ± SE ( n = 8 per group). * P

    Article Snippet: Antibodies were as follows: mouse monoclonal PHB antibody (Thermo Fisher, Fremont, CA), mouse monoclonal HO-1 (Abcam, Cambridge, MA), rabbit polyclonal histone H3 (Millipore, Billerica, MA), mouse monoclonal green fluorescent protein (GFP; Santa Cruz Biotechnology, Santa Cruz, CA), rabbit polyclonal Nrf2 (Santa Cruz Biotechnology), goat polyclonal NQO-1 (Santa Cruz Biotechnology), rabbit polyclonal c-Jun (Santa Cruz Biotechnology), goat polyclonal c-Fos (Santa Cruz Biotechnology), rabbit polyclonal ERK (Santa Cruz Biotechnology), and phosphorylated ERK (Cell Signaling, Danvers, MA).

    Techniques: Mouse Assay, Activity Assay, Quantitative RT-PCR, Isolation

    PHB-induced ARE activation, NQO-1 and HO-1 protein expression, and decreased intracellular reactive oxygen species (ROS) levels during TNFα treatment are not exclusively mediated via Nrf2. A : relative ARE-driven luciferase activity in Caco-2-BBE cells cotransfected with pEGFPN1-PHB (PHB) or pEGFPN1-vector (V) and Nrf2 small interfering RNA (siNrf2) or negative control small interfering RNA (siRNA, TNFα). Values are means ± SE ( n = 6 per treatment). * P

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Nrf2 is not required for epithelial prohibitin-dependent attenuation of experimental colitis

    doi: 10.1152/ajpgi.00327.2012

    Figure Lengend Snippet: PHB-induced ARE activation, NQO-1 and HO-1 protein expression, and decreased intracellular reactive oxygen species (ROS) levels during TNFα treatment are not exclusively mediated via Nrf2. A : relative ARE-driven luciferase activity in Caco-2-BBE cells cotransfected with pEGFPN1-PHB (PHB) or pEGFPN1-vector (V) and Nrf2 small interfering RNA (siNrf2) or negative control small interfering RNA (siRNA, TNFα). Values are means ± SE ( n = 6 per treatment). * P

    Article Snippet: Antibodies were as follows: mouse monoclonal PHB antibody (Thermo Fisher, Fremont, CA), mouse monoclonal HO-1 (Abcam, Cambridge, MA), rabbit polyclonal histone H3 (Millipore, Billerica, MA), mouse monoclonal green fluorescent protein (GFP; Santa Cruz Biotechnology, Santa Cruz, CA), rabbit polyclonal Nrf2 (Santa Cruz Biotechnology), goat polyclonal NQO-1 (Santa Cruz Biotechnology), rabbit polyclonal c-Jun (Santa Cruz Biotechnology), goat polyclonal c-Fos (Santa Cruz Biotechnology), rabbit polyclonal ERK (Santa Cruz Biotechnology), and phosphorylated ERK (Cell Signaling, Danvers, MA).

    Techniques: Activation Assay, Expressing, Luciferase, Activity Assay, Plasmid Preparation, Small Interfering RNA, Negative Control

    Caco-2-BBE cells overexpressing prohibitin (PHB) exhibit increased TNFα-induced nuclear factor erythroid 2-related factor 2 (Nrf2) nuclear localization, antioxidant response element (ARE) activation, and heme oxygenase-1 (HO-1) and NAD(P)H quinone oxidoreductase-1 (NQO-1) expression. A : Western blot of nuclear and cytosolic Nrf2 protein expression in Caco-2-BBE cells overexpressing pEGFPN1-PHB (PHB) or pEGFPN1-vector (V) and treated with 10 ng/ml TNFα for 60 min. Results are representative of 3 independent experiments. Histone H3 and β-actin were used as loading controls for nuclear and cytosolic extracts, respectively. B : relative ARE-driven luciferase activity in Caco-2-BBE cells. Values are means ± SE ( n = 3 per treatment). *** P

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Nrf2 is not required for epithelial prohibitin-dependent attenuation of experimental colitis

    doi: 10.1152/ajpgi.00327.2012

    Figure Lengend Snippet: Caco-2-BBE cells overexpressing prohibitin (PHB) exhibit increased TNFα-induced nuclear factor erythroid 2-related factor 2 (Nrf2) nuclear localization, antioxidant response element (ARE) activation, and heme oxygenase-1 (HO-1) and NAD(P)H quinone oxidoreductase-1 (NQO-1) expression. A : Western blot of nuclear and cytosolic Nrf2 protein expression in Caco-2-BBE cells overexpressing pEGFPN1-PHB (PHB) or pEGFPN1-vector (V) and treated with 10 ng/ml TNFα for 60 min. Results are representative of 3 independent experiments. Histone H3 and β-actin were used as loading controls for nuclear and cytosolic extracts, respectively. B : relative ARE-driven luciferase activity in Caco-2-BBE cells. Values are means ± SE ( n = 3 per treatment). *** P

    Article Snippet: Antibodies were as follows: mouse monoclonal PHB antibody (Thermo Fisher, Fremont, CA), mouse monoclonal HO-1 (Abcam, Cambridge, MA), rabbit polyclonal histone H3 (Millipore, Billerica, MA), mouse monoclonal green fluorescent protein (GFP; Santa Cruz Biotechnology, Santa Cruz, CA), rabbit polyclonal Nrf2 (Santa Cruz Biotechnology), goat polyclonal NQO-1 (Santa Cruz Biotechnology), rabbit polyclonal c-Jun (Santa Cruz Biotechnology), goat polyclonal c-Fos (Santa Cruz Biotechnology), rabbit polyclonal ERK (Santa Cruz Biotechnology), and phosphorylated ERK (Cell Signaling, Danvers, MA).

    Techniques: Activation Assay, Expressing, Western Blot, Plasmid Preparation, Luciferase, Activity Assay

    PHB Tg mice exhibit increased colonic HO-1 and NQO-1 expression during DSS colitis, which is unaffected by Nrf2 deletion. A : qRT-PCR analysis of HO-1, NQO-1, and Nrf2 in total RNA isolated from colon. Values are means ± SE ( n = 8 per group). * P

    Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

    Article Title: Nrf2 is not required for epithelial prohibitin-dependent attenuation of experimental colitis

    doi: 10.1152/ajpgi.00327.2012

    Figure Lengend Snippet: PHB Tg mice exhibit increased colonic HO-1 and NQO-1 expression during DSS colitis, which is unaffected by Nrf2 deletion. A : qRT-PCR analysis of HO-1, NQO-1, and Nrf2 in total RNA isolated from colon. Values are means ± SE ( n = 8 per group). * P

    Article Snippet: Antibodies were as follows: mouse monoclonal PHB antibody (Thermo Fisher, Fremont, CA), mouse monoclonal HO-1 (Abcam, Cambridge, MA), rabbit polyclonal histone H3 (Millipore, Billerica, MA), mouse monoclonal green fluorescent protein (GFP; Santa Cruz Biotechnology, Santa Cruz, CA), rabbit polyclonal Nrf2 (Santa Cruz Biotechnology), goat polyclonal NQO-1 (Santa Cruz Biotechnology), rabbit polyclonal c-Jun (Santa Cruz Biotechnology), goat polyclonal c-Fos (Santa Cruz Biotechnology), rabbit polyclonal ERK (Santa Cruz Biotechnology), and phosphorylated ERK (Cell Signaling, Danvers, MA).

    Techniques: Mouse Assay, Expressing, Quantitative RT-PCR, Isolation

    Comparison of Hmox1 protein expression in mouse and human liver tissue of controls with Hmox1-deficient animals and patient samples. (A) Cross-sections of paraffin-embedded liver tissue immunofluorescence showed that Kupffer cells were a major site of Hmox1 expression in both WT Ctr and WT BMT mice (left); no specific Hmox1 signal was detectable in the liver KO Ctr animals (upper right); an Hmox1-expressing Kupffer cell population was restored in the liver of transplant recipients (bottom right). Arrows point to Hmox1-positive cells. (B) A human liver biopsy specimen obtained from human with intact HMOX1 gene showed high HMOX1 signal present in Kupffer-like cells (left), whereas there was no HMOX1 expression in the liver sample of an HMOX1-deficient patient specimen (right). Scale bars represent 50 μm (A) and 25 μm (B).

    Journal: Blood

    Article Title: Wild-type macrophages reverse disease in heme oxygenase 1-deficient mice

    doi: 10.1182/blood-2014-02-554162

    Figure Lengend Snippet: Comparison of Hmox1 protein expression in mouse and human liver tissue of controls with Hmox1-deficient animals and patient samples. (A) Cross-sections of paraffin-embedded liver tissue immunofluorescence showed that Kupffer cells were a major site of Hmox1 expression in both WT Ctr and WT BMT mice (left); no specific Hmox1 signal was detectable in the liver KO Ctr animals (upper right); an Hmox1-expressing Kupffer cell population was restored in the liver of transplant recipients (bottom right). Arrows point to Hmox1-positive cells. (B) A human liver biopsy specimen obtained from human with intact HMOX1 gene showed high HMOX1 signal present in Kupffer-like cells (left), whereas there was no HMOX1 expression in the liver sample of an HMOX1-deficient patient specimen (right). Scale bars represent 50 μm (A) and 25 μm (B).

    Article Snippet: Hmox1 protein expression in mouse liver was evaluated with monoclonal mouse anti-Hmox1 primary (ab13248; Abcam) and fluorescein isothiocyanate-labeled goat anti-mouse secondary antibodies.

    Techniques: Expressing, Immunofluorescence, Mouse Assay

    BMT reduced the oxidative stress response and prevented injury to Hmox1 −/− kidneys. mRNA levels of oxidative stress responsive genes were significantly elevated in KO Ctr animals, including Gsta2 (glutamine- S -transferase A2) (A), Gstm1 (glutamine- S -transferase μ 1) (B), Gclc (glutamate-cysteine ligase, catalytic subunit) (C), Nqo1 [NAD(P)H:quinone dehydrogenase, quinone 1] (D), Fpn1 (ferroportin 1) (E), and Mrp2 (F), but returned to normal after BMT. (G) Masson’s trichrome staining of paraffin-embedded kidney tissues revealed significant accumulation of collagen, which is colored blue (arrows) in KO Ctr mice. Collagen depositions were minimal in transplanted animals. Scale bar represents 50 μm.

    Journal: Blood

    Article Title: Wild-type macrophages reverse disease in heme oxygenase 1-deficient mice

    doi: 10.1182/blood-2014-02-554162

    Figure Lengend Snippet: BMT reduced the oxidative stress response and prevented injury to Hmox1 −/− kidneys. mRNA levels of oxidative stress responsive genes were significantly elevated in KO Ctr animals, including Gsta2 (glutamine- S -transferase A2) (A), Gstm1 (glutamine- S -transferase μ 1) (B), Gclc (glutamate-cysteine ligase, catalytic subunit) (C), Nqo1 [NAD(P)H:quinone dehydrogenase, quinone 1] (D), Fpn1 (ferroportin 1) (E), and Mrp2 (F), but returned to normal after BMT. (G) Masson’s trichrome staining of paraffin-embedded kidney tissues revealed significant accumulation of collagen, which is colored blue (arrows) in KO Ctr mice. Collagen depositions were minimal in transplanted animals. Scale bar represents 50 μm.

    Article Snippet: Hmox1 protein expression in mouse liver was evaluated with monoclonal mouse anti-Hmox1 primary (ab13248; Abcam) and fluorescein isothiocyanate-labeled goat anti-mouse secondary antibodies.

    Techniques: Staining, Mouse Assay

    Hmox1 mRNA expression and quantification of Hmox1 +/+ DNA in transplanted Hmox1 −/− animals. Hmox1 mRNA expression levels in the tissues of KO BMT animals were partially restored to normal in the BM (A) and spleen (B), were twofold higher in the liver (C), and were negligibly low in the kidney (D) in comparison with WT animals. Data were obtained by quantitative reverse-transcription polymerase chain reaction. ND, not detected. (E) Percentage of WT cells in the tissues of KO BMT animals, as assessed by quantification of the WT Hmox1 gene in a total genomic DNA extracts; we observed that liver had the highest number of donor cells of the 3 key tissues analyzed, at 3%. Primers that target exon 3 of the Hmox1 gene, which was deleted in Hmox1 −/− mice, were used for the quantification.

    Journal: Blood

    Article Title: Wild-type macrophages reverse disease in heme oxygenase 1-deficient mice

    doi: 10.1182/blood-2014-02-554162

    Figure Lengend Snippet: Hmox1 mRNA expression and quantification of Hmox1 +/+ DNA in transplanted Hmox1 −/− animals. Hmox1 mRNA expression levels in the tissues of KO BMT animals were partially restored to normal in the BM (A) and spleen (B), were twofold higher in the liver (C), and were negligibly low in the kidney (D) in comparison with WT animals. Data were obtained by quantitative reverse-transcription polymerase chain reaction. ND, not detected. (E) Percentage of WT cells in the tissues of KO BMT animals, as assessed by quantification of the WT Hmox1 gene in a total genomic DNA extracts; we observed that liver had the highest number of donor cells of the 3 key tissues analyzed, at 3%. Primers that target exon 3 of the Hmox1 gene, which was deleted in Hmox1 −/− mice, were used for the quantification.

    Article Snippet: Hmox1 protein expression in mouse liver was evaluated with monoclonal mouse anti-Hmox1 primary (ab13248; Abcam) and fluorescein isothiocyanate-labeled goat anti-mouse secondary antibodies.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Mouse Assay

    Kupffer cells are absent in the livers of Hmox1 −/− animals and a human patient, but BMT restores Kupffer cells to Hmox1 −/− mice. (A) Immunohistochemistry for the pan-macrophage marker, F4/80, indicated that Kupffer cells were virtually absent in the livers of Hmox1 −/− sham mice (upper middle and right) in comparison with WT mice (left). F4/80-positive cells turned brown after diaminobenzidine staining (arrowheads). BMT completely restored macrophage populations in Hmox1 −/− animals (lower middle and right). Results are shown for Hmox1 −/− animals that were 4 months (middle) or 1.7 months (right) old at the time the BMT procedure was performed. (B) Expression of the marker of M2 polarized macrophages indicated that CD163 was partially restored in liver, BM, and spleen of KO BMT mice, as determined by quantitative reverse-transcription polymerase chain reaction. ND, not detected. (C) CD163 Kupffer cells normally found in human liver (left) were undetectable in the liver of an HMOX1-deficient patient (right). Scale bars represent 50 μm (A) and 25 μm (C).

    Journal: Blood

    Article Title: Wild-type macrophages reverse disease in heme oxygenase 1-deficient mice

    doi: 10.1182/blood-2014-02-554162

    Figure Lengend Snippet: Kupffer cells are absent in the livers of Hmox1 −/− animals and a human patient, but BMT restores Kupffer cells to Hmox1 −/− mice. (A) Immunohistochemistry for the pan-macrophage marker, F4/80, indicated that Kupffer cells were virtually absent in the livers of Hmox1 −/− sham mice (upper middle and right) in comparison with WT mice (left). F4/80-positive cells turned brown after diaminobenzidine staining (arrowheads). BMT completely restored macrophage populations in Hmox1 −/− animals (lower middle and right). Results are shown for Hmox1 −/− animals that were 4 months (middle) or 1.7 months (right) old at the time the BMT procedure was performed. (B) Expression of the marker of M2 polarized macrophages indicated that CD163 was partially restored in liver, BM, and spleen of KO BMT mice, as determined by quantitative reverse-transcription polymerase chain reaction. ND, not detected. (C) CD163 Kupffer cells normally found in human liver (left) were undetectable in the liver of an HMOX1-deficient patient (right). Scale bars represent 50 μm (A) and 25 μm (C).

    Article Snippet: Hmox1 protein expression in mouse liver was evaluated with monoclonal mouse anti-Hmox1 primary (ab13248; Abcam) and fluorescein isothiocyanate-labeled goat anti-mouse secondary antibodies.

    Techniques: Mouse Assay, Immunohistochemistry, Marker, Staining, Expressing, Reverse Transcription Polymerase Chain Reaction

    Nonmyeloablative BM transplantation in Hmox1 −/− mice improved blood chemistries and led to resolution of anemia. (A) Scheme of the BM transplantation experiment used for subsequent data collection. (B) BM engraftment dynamics are represented as a percentage of CD45.2-positive leukocytes in peripheral blood estimated by fluorescence-activated cell sorter analysis. Changes over weeks are plotted for each individual mouse. (C) Hmox1 −/− mice had elevated serum ALP and LDH levels. The blood chemistries returned to normal in BM transplanted recipients. (D) Indicators of microcytic anemia mean cell volume and hematocrit improved in BM-transplanted Hmox1 −/− mice. (C-D) Average values for the terminal time, 21 weeks, are shown for each experimental group; error bars represent the standard deviation (N = 5). TMS water, drinking water supplemented with trimethoprim (300 μg/ml) and sulfamethoxazole (60 μg/ml).

    Journal: Blood

    Article Title: Wild-type macrophages reverse disease in heme oxygenase 1-deficient mice

    doi: 10.1182/blood-2014-02-554162

    Figure Lengend Snippet: Nonmyeloablative BM transplantation in Hmox1 −/− mice improved blood chemistries and led to resolution of anemia. (A) Scheme of the BM transplantation experiment used for subsequent data collection. (B) BM engraftment dynamics are represented as a percentage of CD45.2-positive leukocytes in peripheral blood estimated by fluorescence-activated cell sorter analysis. Changes over weeks are plotted for each individual mouse. (C) Hmox1 −/− mice had elevated serum ALP and LDH levels. The blood chemistries returned to normal in BM transplanted recipients. (D) Indicators of microcytic anemia mean cell volume and hematocrit improved in BM-transplanted Hmox1 −/− mice. (C-D) Average values for the terminal time, 21 weeks, are shown for each experimental group; error bars represent the standard deviation (N = 5). TMS water, drinking water supplemented with trimethoprim (300 μg/ml) and sulfamethoxazole (60 μg/ml).

    Article Snippet: Hmox1 protein expression in mouse liver was evaluated with monoclonal mouse anti-Hmox1 primary (ab13248; Abcam) and fluorescein isothiocyanate-labeled goat anti-mouse secondary antibodies.

    Techniques: Transplantation Assay, Mouse Assay, Fluorescence, ALP Assay, Standard Deviation

    Effect of HO-1 on neurologic deficits, brain injury volume, and brain edema. (a) Neurologic deficits of mice after collagenase-induced ICH; n = 8 mice/group. (b) Brain injury volume on day 3 after collagenase-induced ICH; n = 8 mice/group. (c) Brain water content of mice on day 3 after collagenase-induced ICH. Ipsi-Stri, ipsilateral striatum; Con-Stri, contralateral striatum; Cerebel, cerebellum. (d) Hemoglobin content in ipsilateral striatal tissue of brains was determined by Drabkin’s reagent on day 1 day after ICH. n = 5 mice/group. * p

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: Distinct role of heme oxygenase-1 in early- and late-stage intracerebral hemorrhage in 12-month-old mice

    doi: 10.1177/0271678X16655814

    Figure Lengend Snippet: Effect of HO-1 on neurologic deficits, brain injury volume, and brain edema. (a) Neurologic deficits of mice after collagenase-induced ICH; n = 8 mice/group. (b) Brain injury volume on day 3 after collagenase-induced ICH; n = 8 mice/group. (c) Brain water content of mice on day 3 after collagenase-induced ICH. Ipsi-Stri, ipsilateral striatum; Con-Stri, contralateral striatum; Cerebel, cerebellum. (d) Hemoglobin content in ipsilateral striatal tissue of brains was determined by Drabkin’s reagent on day 1 day after ICH. n = 5 mice/group. * p

    Article Snippet: Western blot analysis was carried out as in our previous study ( n = 5 mice/group)., The primary antibodies used were mouse monoclonal anti-HO-1 (1:1000; Stressgen), rabbit anti-ZO-1 (1:200; Abcam), rabbit anti-occludin (1:500; Abcam), rabbit anti-MMP-3 (1:500; Abcam), and mouse monoclonal anti-β-actin (1: 3000; Abcam).

    Techniques: Mouse Assay

    Effect of HO-1 on hematoma clearance and angiogenesis. (a) Hematoma volume was assessed by MRI. Hematoma was detected in T1-weighted images as a low signal on days 1 and 3 after ICH and then as a high signal on day 7. Residual hematoma was observed as a low signal on days 14 and 28 after ICH. n = 5 mice/group. * p

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: Distinct role of heme oxygenase-1 in early- and late-stage intracerebral hemorrhage in 12-month-old mice

    doi: 10.1177/0271678X16655814

    Figure Lengend Snippet: Effect of HO-1 on hematoma clearance and angiogenesis. (a) Hematoma volume was assessed by MRI. Hematoma was detected in T1-weighted images as a low signal on days 1 and 3 after ICH and then as a high signal on day 7. Residual hematoma was observed as a low signal on days 14 and 28 after ICH. n = 5 mice/group. * p

    Article Snippet: Western blot analysis was carried out as in our previous study ( n = 5 mice/group)., The primary antibodies used were mouse monoclonal anti-HO-1 (1:1000; Stressgen), rabbit anti-ZO-1 (1:200; Abcam), rabbit anti-occludin (1:500; Abcam), rabbit anti-MMP-3 (1:500; Abcam), and mouse monoclonal anti-β-actin (1: 3000; Abcam).

    Techniques: Magnetic Resonance Imaging, Mouse Assay

    Identification and expression of HO-1. (a) HO-1 expression in normal brain tissue. Scale bars = 50 µm, n = 3 mice/group. (b) Identification of HO-1 in the perihematomal region on day 3 after collagenase-induced ICH. Scale bars = 50 µm, n = 5 mice/group. Arrows indicate cells with colocalization. Insets represent higher magnification of cells with colocalization. (c) HO-1 protein level in ipsilateral brain after collagenase-induced ICH. (d) Quantification of HO-1 protein level after collagenase-induced ICH. Ipsi-Stri, ipsilateral striatum; Con-Stri, contralateral striatum. n = 5 mice/group. Values are mean ± SD. # p

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: Distinct role of heme oxygenase-1 in early- and late-stage intracerebral hemorrhage in 12-month-old mice

    doi: 10.1177/0271678X16655814

    Figure Lengend Snippet: Identification and expression of HO-1. (a) HO-1 expression in normal brain tissue. Scale bars = 50 µm, n = 3 mice/group. (b) Identification of HO-1 in the perihematomal region on day 3 after collagenase-induced ICH. Scale bars = 50 µm, n = 5 mice/group. Arrows indicate cells with colocalization. Insets represent higher magnification of cells with colocalization. (c) HO-1 protein level in ipsilateral brain after collagenase-induced ICH. (d) Quantification of HO-1 protein level after collagenase-induced ICH. Ipsi-Stri, ipsilateral striatum; Con-Stri, contralateral striatum. n = 5 mice/group. Values are mean ± SD. # p

    Article Snippet: Western blot analysis was carried out as in our previous study ( n = 5 mice/group)., The primary antibodies used were mouse monoclonal anti-HO-1 (1:1000; Stressgen), rabbit anti-ZO-1 (1:200; Abcam), rabbit anti-occludin (1:500; Abcam), rabbit anti-MMP-3 (1:500; Abcam), and mouse monoclonal anti-β-actin (1: 3000; Abcam).

    Techniques: Expressing, Mouse Assay

    Effect of HO-1 induction and inhibition on reactive oxygen species (ROS) production, superoxide dismutase (SOD) activity, and malondialdehyde (MDA) content in hemin-exposed BV2 microglia. (a, b) ROS production in BV2 microglia. The Bv2 microglia were labeled by anti-Iba1 antibody (green), and ROS production was detected by dihydroethidine (red). Values are means ± SD. # p

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: Distinct role of heme oxygenase-1 in early- and late-stage intracerebral hemorrhage in 12-month-old mice

    doi: 10.1177/0271678X16655814

    Figure Lengend Snippet: Effect of HO-1 induction and inhibition on reactive oxygen species (ROS) production, superoxide dismutase (SOD) activity, and malondialdehyde (MDA) content in hemin-exposed BV2 microglia. (a, b) ROS production in BV2 microglia. The Bv2 microglia were labeled by anti-Iba1 antibody (green), and ROS production was detected by dihydroethidine (red). Values are means ± SD. # p

    Article Snippet: Western blot analysis was carried out as in our previous study ( n = 5 mice/group)., The primary antibodies used were mouse monoclonal anti-HO-1 (1:1000; Stressgen), rabbit anti-ZO-1 (1:200; Abcam), rabbit anti-occludin (1:500; Abcam), rabbit anti-MMP-3 (1:500; Abcam), and mouse monoclonal anti-β-actin (1: 3000; Abcam).

    Techniques: Inhibition, Activity Assay, Multiple Displacement Amplification, Labeling

    Effect of HO-1 on oxidative damage. (a) Reactive oxygen species (ROS) signal fluorescence intensity in the perihematomal region on day 3 after collagenase-induced ICH. n = 8 mice/group. (b) The expression of 4-HNE in the perihematomal region on day 3 after collagenase-induced ICH. n = 5 mice/group. * p

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: Distinct role of heme oxygenase-1 in early- and late-stage intracerebral hemorrhage in 12-month-old mice

    doi: 10.1177/0271678X16655814

    Figure Lengend Snippet: Effect of HO-1 on oxidative damage. (a) Reactive oxygen species (ROS) signal fluorescence intensity in the perihematomal region on day 3 after collagenase-induced ICH. n = 8 mice/group. (b) The expression of 4-HNE in the perihematomal region on day 3 after collagenase-induced ICH. n = 5 mice/group. * p

    Article Snippet: Western blot analysis was carried out as in our previous study ( n = 5 mice/group)., The primary antibodies used were mouse monoclonal anti-HO-1 (1:1000; Stressgen), rabbit anti-ZO-1 (1:200; Abcam), rabbit anti-occludin (1:500; Abcam), rabbit anti-MMP-3 (1:500; Abcam), and mouse monoclonal anti-β-actin (1: 3000; Abcam).

    Techniques: Fluorescence, Mouse Assay, Expressing

    Effect of HO-1 on blood–brain barrier after collagenase-induced ICH. (a) Evans blue (EB) extravasation on day 3 after ICH. (b, c) Western blot analysis of tight junction proteins ZO-1 (b) and occludin (c) on day 3 after collagenase-induced ICH. (d) Gelatin gel zymography analysis shows activation of matrix metalloproteinase (MMP)-9 and MMP-2 on day 3 after ICH. n = 5 mice/group, * p

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: Distinct role of heme oxygenase-1 in early- and late-stage intracerebral hemorrhage in 12-month-old mice

    doi: 10.1177/0271678X16655814

    Figure Lengend Snippet: Effect of HO-1 on blood–brain barrier after collagenase-induced ICH. (a) Evans blue (EB) extravasation on day 3 after ICH. (b, c) Western blot analysis of tight junction proteins ZO-1 (b) and occludin (c) on day 3 after collagenase-induced ICH. (d) Gelatin gel zymography analysis shows activation of matrix metalloproteinase (MMP)-9 and MMP-2 on day 3 after ICH. n = 5 mice/group, * p

    Article Snippet: Western blot analysis was carried out as in our previous study ( n = 5 mice/group)., The primary antibodies used were mouse monoclonal anti-HO-1 (1:1000; Stressgen), rabbit anti-ZO-1 (1:200; Abcam), rabbit anti-occludin (1:500; Abcam), rabbit anti-MMP-3 (1:500; Abcam), and mouse monoclonal anti-β-actin (1: 3000; Abcam).

    Techniques: Western Blot, Zymography, Activation Assay, Mouse Assay

    Effect of HO-1 on inflammation and white matter injury. (a) The expression of Iba1-, glial fibrillary acidic protein (GFAP)-, and myeloperoxidase (MPO)-immunoreactive cells around the injury site on day 3 after collagenase-induced ICH; n = 8 mice/group. The bar graph shows quantification analysis. (b) Immunofluorescent and Luxol fast blue (LFB) staining showing the expression of degraded myelin basic protein (dMBP), amyloid precursor protein (APP), and normal myelin (LFB) in the perihematoma region on day 3 after ICH. Arrows indicate damaged myelin, damaged axons, and normal myelin. The bar graphs show quantification analysis. n = 8 mice/group. * p

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: Distinct role of heme oxygenase-1 in early- and late-stage intracerebral hemorrhage in 12-month-old mice

    doi: 10.1177/0271678X16655814

    Figure Lengend Snippet: Effect of HO-1 on inflammation and white matter injury. (a) The expression of Iba1-, glial fibrillary acidic protein (GFAP)-, and myeloperoxidase (MPO)-immunoreactive cells around the injury site on day 3 after collagenase-induced ICH; n = 8 mice/group. The bar graph shows quantification analysis. (b) Immunofluorescent and Luxol fast blue (LFB) staining showing the expression of degraded myelin basic protein (dMBP), amyloid precursor protein (APP), and normal myelin (LFB) in the perihematoma region on day 3 after ICH. Arrows indicate damaged myelin, damaged axons, and normal myelin. The bar graphs show quantification analysis. n = 8 mice/group. * p

    Article Snippet: Western blot analysis was carried out as in our previous study ( n = 5 mice/group)., The primary antibodies used were mouse monoclonal anti-HO-1 (1:1000; Stressgen), rabbit anti-ZO-1 (1:200; Abcam), rabbit anti-occludin (1:500; Abcam), rabbit anti-MMP-3 (1:500; Abcam), and mouse monoclonal anti-β-actin (1: 3000; Abcam).

    Techniques: Expressing, Mouse Assay, Staining

    TQ increased Nrf2, HO-1 and SOD inhibition rate in the cortex and hippocampus of SE rats. (A) Protein extracted from hippocampal and cortical tissues was assayed by western blotting. Nrf2, HO-1 expression in SE rat brains was significantly lower than in control group rats. TQ significantly downregulated the expression of Nrf2 and HO-1. (B) The inhibition rate of SOD. * Model group vs. control group; ΔTQ group vs. model group, p

    Journal: Translational Neuroscience

    Article Title: Thymoquinone Attenuates Brain Injury via an Anti-oxidative Pathway in a Status Epilepticus Rat Model

    doi: 10.1515/tnsci-2017-0003

    Figure Lengend Snippet: TQ increased Nrf2, HO-1 and SOD inhibition rate in the cortex and hippocampus of SE rats. (A) Protein extracted from hippocampal and cortical tissues was assayed by western blotting. Nrf2, HO-1 expression in SE rat brains was significantly lower than in control group rats. TQ significantly downregulated the expression of Nrf2 and HO-1. (B) The inhibition rate of SOD. * Model group vs. control group; ΔTQ group vs. model group, p

    Article Snippet: The membrane was incubated with anti-Nrf2 rabbit polyclonal antibody (1:500; Santa Cruz Biotechnology, USA), anti-HO-1 mouse monoclonal antibody (1:500; Abcam, USA), and anti-alpha tublin primary antibody (1:5000; Proteintech group, USA) at 4°C overnight.

    Techniques: Inhibition, Western Blot, Expressing

    Effect of Nrf2 knockdown on Nrf2 and HO-1 in B16-F10 melanoma cells that were induced by ionizing radiation. B16-F10 melanoma cells were radiated by a dose of 8 Gy for 12 h following the treatment of melanoma cells with Nrf2 siRNA. The cells were harvested and western blotting was performed to detect the expression of (A) Nrf2 protein. (B) Quantitative analysis of Nrf2 protein expression. (C) Western blot analysis of HO-1 protein expression. (D) Quantitative analysis of HO-1 protein expression. (E) Reverse transcription-quantitative polymerase chain reaction of Nrf2 mRNA. (F) Analysis of HO-1 activity. All data are expressed as the mean ± standard deviation *P

    Journal: Oncology Letters

    Article Title: Migration and invasion in B16-F10 mouse melanoma cells are regulated by Nrf2 inhibition during treatment with ionizing radiation

    doi: 10.3892/ol.2018.8799

    Figure Lengend Snippet: Effect of Nrf2 knockdown on Nrf2 and HO-1 in B16-F10 melanoma cells that were induced by ionizing radiation. B16-F10 melanoma cells were radiated by a dose of 8 Gy for 12 h following the treatment of melanoma cells with Nrf2 siRNA. The cells were harvested and western blotting was performed to detect the expression of (A) Nrf2 protein. (B) Quantitative analysis of Nrf2 protein expression. (C) Western blot analysis of HO-1 protein expression. (D) Quantitative analysis of HO-1 protein expression. (E) Reverse transcription-quantitative polymerase chain reaction of Nrf2 mRNA. (F) Analysis of HO-1 activity. All data are expressed as the mean ± standard deviation *P

    Article Snippet: The membranes were blocked with blocking solution [0.05% Tween and 5% bovine serum albumin (BSA; BBI Life Sciences Corp., Shanghai, China)] in Tris-buffered saline for 2 h at room temperature and incubated with primary rabbit monoclonal antibodies against Nrf2 (cat. no. ab62352; 1:500), primary mouse monoclonal antibodies against HO-1 (cat. no. ab13248; 1:500), primary rabbit polyclonal antibodies against cleaved caspase-3 (cat. no. ab2302; 1:500), or primary mouse monoclonal antibodies against β-actin (cat. no. ab8226; 1:1,000; all Abcam, Cambridge UK), overnight at 4°C.

    Techniques: Western Blot, Expressing, Real-time Polymerase Chain Reaction, Activity Assay, Standard Deviation

    Expression of haem oxygenase 1 (HO-1) is reduced in FcγRIIb-deficient mice. Single cell suspensions of splenocytes from FcγRIIb-deficient and C57BL/6 wild type (WT) mice at 12 months were obtained. (a) Total mRNA was purified and

    Journal: Immunology

    Article Title: Carbon monoxide exposure improves immune function in lupus-prone mice

    doi: 10.1111/imm.12124

    Figure Lengend Snippet: Expression of haem oxygenase 1 (HO-1) is reduced in FcγRIIb-deficient mice. Single cell suspensions of splenocytes from FcγRIIb-deficient and C57BL/6 wild type (WT) mice at 12 months were obtained. (a) Total mRNA was purified and

    Article Snippet: FITC-conjugated anti-mouse HO-1 monoclonal antibody (clone HO-1-2) and anti-mouse HO-1 monoclonal antibody (clone HO-1-1) were purchased from Abcam (Cambridge, UK).

    Techniques: Expressing, Mouse Assay, Purification

    Effects of nitrogen mustard on HO-1 expression in rabbit corneas

    Journal: Toxicology and applied pharmacology

    Article Title: The generation of 4-hydroxynonenal, an electrophilic lipid peroxidation end product, in rabbit cornea organ cultures treated with UVB light and nitrogen mustard

    doi: 10.1016/j.taap.2013.06.025

    Figure Lengend Snippet: Effects of nitrogen mustard on HO-1 expression in rabbit corneas

    Article Snippet: Mouse monoclonal 4-HNE antibody was from R & D Systems (Minneapolis, MN), rabbit anti-HO-1 polyclonal antibody from Stressgen Biotechnology (Victoria, BC, Canada) and mouse monoclonal anti-HO-1 antibody from Abcam (Cambridge, MA).

    Techniques: Expressing

    Effects of electrophilic lipid peroxidation products on HO-1 expression in human corneal epithelial cells

    Journal: Toxicology and applied pharmacology

    Article Title: The generation of 4-hydroxynonenal, an electrophilic lipid peroxidation end product, in rabbit cornea organ cultures treated with UVB light and nitrogen mustard

    doi: 10.1016/j.taap.2013.06.025

    Figure Lengend Snippet: Effects of electrophilic lipid peroxidation products on HO-1 expression in human corneal epithelial cells

    Article Snippet: Mouse monoclonal 4-HNE antibody was from R & D Systems (Minneapolis, MN), rabbit anti-HO-1 polyclonal antibody from Stressgen Biotechnology (Victoria, BC, Canada) and mouse monoclonal anti-HO-1 antibody from Abcam (Cambridge, MA).

    Techniques: Expressing

    Role of MAP kinases and PI3/Akt kinase signaling in 4-HNE-induced HO-1 expression in human corneal epithelial cells

    Journal: Toxicology and applied pharmacology

    Article Title: The generation of 4-hydroxynonenal, an electrophilic lipid peroxidation end product, in rabbit cornea organ cultures treated with UVB light and nitrogen mustard

    doi: 10.1016/j.taap.2013.06.025

    Figure Lengend Snippet: Role of MAP kinases and PI3/Akt kinase signaling in 4-HNE-induced HO-1 expression in human corneal epithelial cells

    Article Snippet: Mouse monoclonal 4-HNE antibody was from R & D Systems (Minneapolis, MN), rabbit anti-HO-1 polyclonal antibody from Stressgen Biotechnology (Victoria, BC, Canada) and mouse monoclonal anti-HO-1 antibody from Abcam (Cambridge, MA).

    Techniques: Expressing

    Effects of UVB on HO-1 expression in rabbit corneas

    Journal: Toxicology and applied pharmacology

    Article Title: The generation of 4-hydroxynonenal, an electrophilic lipid peroxidation end product, in rabbit cornea organ cultures treated with UVB light and nitrogen mustard

    doi: 10.1016/j.taap.2013.06.025

    Figure Lengend Snippet: Effects of UVB on HO-1 expression in rabbit corneas

    Article Snippet: Mouse monoclonal 4-HNE antibody was from R & D Systems (Minneapolis, MN), rabbit anti-HO-1 polyclonal antibody from Stressgen Biotechnology (Victoria, BC, Canada) and mouse monoclonal anti-HO-1 antibody from Abcam (Cambridge, MA).

    Techniques: Expressing

    HO-1 metabolites mediate resistance effects to chemotherapy . Cell growth was assessed 72 h after application of gemcitabine via MTT assay. (5A) Biliverdin administration biliverdin [5 and 50 μmol/l] resulted in increased proliferation of cancer cell line PANC-1 and S2-013, and therefore reduced the chemotherapeutic effect of gemcitabine (LD50 dose). (5B) Administration of ferrous histidinate revealed increased cell proliferation of PANC-1 and S2-013 cancer cells.

    Journal: Molecular Cancer

    Article Title: Heme oxygenase-1 and its metabolites affect pancreatic tumor growth in vivo

    doi: 10.1186/1476-4598-8-37

    Figure Lengend Snippet: HO-1 metabolites mediate resistance effects to chemotherapy . Cell growth was assessed 72 h after application of gemcitabine via MTT assay. (5A) Biliverdin administration biliverdin [5 and 50 μmol/l] resulted in increased proliferation of cancer cell line PANC-1 and S2-013, and therefore reduced the chemotherapeutic effect of gemcitabine (LD50 dose). (5B) Administration of ferrous histidinate revealed increased cell proliferation of PANC-1 and S2-013 cancer cells.

    Article Snippet: The membranes were probed with primary anti-HO-1 mouse monoclonal antibody (BD Biosciences Transduction Laboratories, Lexington, KY, USA).

    Techniques: MTT Assay

    Different HO-1 expression levels are associated with variable susceptibility to chemotherapy . Cell viability was assessed 72 h after application of gemcitabine via MTT assay. Both cell lines showed dose dependent growth inhibition under treatment with gemcitabine. S2-013 cell line with low native HO-1 expression was significantly more susceptible to gemcitabine (LD50 [S2-013] ~2.13 ng/ml) than PANC-1 cell line with high native HO-1 expression (LD50 [PANC-1] ~7.53 ng/ml).

    Journal: Molecular Cancer

    Article Title: Heme oxygenase-1 and its metabolites affect pancreatic tumor growth in vivo

    doi: 10.1186/1476-4598-8-37

    Figure Lengend Snippet: Different HO-1 expression levels are associated with variable susceptibility to chemotherapy . Cell viability was assessed 72 h after application of gemcitabine via MTT assay. Both cell lines showed dose dependent growth inhibition under treatment with gemcitabine. S2-013 cell line with low native HO-1 expression was significantly more susceptible to gemcitabine (LD50 [S2-013] ~2.13 ng/ml) than PANC-1 cell line with high native HO-1 expression (LD50 [PANC-1] ~7.53 ng/ml).

    Article Snippet: The membranes were probed with primary anti-HO-1 mouse monoclonal antibody (BD Biosciences Transduction Laboratories, Lexington, KY, USA).

    Techniques: Expressing, MTT Assay, Inhibition

    Inhibition of HO-1 activity leads to increased susceptibility to chemotherapy in vitro . Cell viability was assessed 72 h after application of gemcitabine via MTT assay. (3A) Under gemcitabine treatment (LD50 dose) application of ZnPP revealed a dose dependent growth inhibition of PANC-1 cancer cells, whereas application of CoPP had no significant effect on cell proliferation. (3B) In S2-013 cell line implementation of CoPP led to increased cell proliferation, whereas ZnPP had no significant effect.

    Journal: Molecular Cancer

    Article Title: Heme oxygenase-1 and its metabolites affect pancreatic tumor growth in vivo

    doi: 10.1186/1476-4598-8-37

    Figure Lengend Snippet: Inhibition of HO-1 activity leads to increased susceptibility to chemotherapy in vitro . Cell viability was assessed 72 h after application of gemcitabine via MTT assay. (3A) Under gemcitabine treatment (LD50 dose) application of ZnPP revealed a dose dependent growth inhibition of PANC-1 cancer cells, whereas application of CoPP had no significant effect on cell proliferation. (3B) In S2-013 cell line implementation of CoPP led to increased cell proliferation, whereas ZnPP had no significant effect.

    Article Snippet: The membranes were probed with primary anti-HO-1 mouse monoclonal antibody (BD Biosciences Transduction Laboratories, Lexington, KY, USA).

    Techniques: Inhibition, Activity Assay, In Vitro, MTT Assay

    Inhibition of HO-1 activity leads to increased susceptibility to chemotherapy of solid tumor growth . Primary pancreatic tumor weights were assessed 4 and 8 weeks after orthotopic tumor implantation of cancer cells in nude mice (n = 60, 15 per group). (4A) ZnPP treatment reduced tumor growth in gemcitabine treated mice after PANC-1 cancer cell implantation significantly, compared to gemcitabine treated tumors only. (4B) Administration of gemcitabine in combination with CoPP or ZnPP in mice implanted S2-013 cancer cells did not significantly change tumor weights in comparison to gemcitabine treatment alone. The use of ZnPP in combination with gemcitabine showed a trend towards lower tumor growth.

    Journal: Molecular Cancer

    Article Title: Heme oxygenase-1 and its metabolites affect pancreatic tumor growth in vivo

    doi: 10.1186/1476-4598-8-37

    Figure Lengend Snippet: Inhibition of HO-1 activity leads to increased susceptibility to chemotherapy of solid tumor growth . Primary pancreatic tumor weights were assessed 4 and 8 weeks after orthotopic tumor implantation of cancer cells in nude mice (n = 60, 15 per group). (4A) ZnPP treatment reduced tumor growth in gemcitabine treated mice after PANC-1 cancer cell implantation significantly, compared to gemcitabine treated tumors only. (4B) Administration of gemcitabine in combination with CoPP or ZnPP in mice implanted S2-013 cancer cells did not significantly change tumor weights in comparison to gemcitabine treatment alone. The use of ZnPP in combination with gemcitabine showed a trend towards lower tumor growth.

    Article Snippet: The membranes were probed with primary anti-HO-1 mouse monoclonal antibody (BD Biosciences Transduction Laboratories, Lexington, KY, USA).

    Techniques: Inhibition, Activity Assay, Tumor Implantation, Mouse Assay

    Pancreatic cancer cell lines PANC-1 and S2-013 show divergent HO-1 expression levels . mRNA extractions from cells for QRT-PCR and cell lysates for Western blot were prepared from untreated cells and cells treated with gemcitabine. (1A) No treatment of PANC-1 cells displayed high native mRNA expression of HO-1, whereas HO-1 expression was only at levels of detection in S2-013. Treatment with gemcitabine increased mRNA expression of HO-1 in PANC-1 cells but did not alter the expression level in S2-013 cancer cells. (1B) Protein analyses in resting condition revealed high native expression of HO-1 in PANC-1 but faint expression in S2-013. Under gemcitabine conditions protein expression of HO-1 was slightly induced in PANC-1 cell line but did not alter HO-1 protein expression in S2-013 cell line.

    Journal: Molecular Cancer

    Article Title: Heme oxygenase-1 and its metabolites affect pancreatic tumor growth in vivo

    doi: 10.1186/1476-4598-8-37

    Figure Lengend Snippet: Pancreatic cancer cell lines PANC-1 and S2-013 show divergent HO-1 expression levels . mRNA extractions from cells for QRT-PCR and cell lysates for Western blot were prepared from untreated cells and cells treated with gemcitabine. (1A) No treatment of PANC-1 cells displayed high native mRNA expression of HO-1, whereas HO-1 expression was only at levels of detection in S2-013. Treatment with gemcitabine increased mRNA expression of HO-1 in PANC-1 cells but did not alter the expression level in S2-013 cancer cells. (1B) Protein analyses in resting condition revealed high native expression of HO-1 in PANC-1 but faint expression in S2-013. Under gemcitabine conditions protein expression of HO-1 was slightly induced in PANC-1 cell line but did not alter HO-1 protein expression in S2-013 cell line.

    Article Snippet: The membranes were probed with primary anti-HO-1 mouse monoclonal antibody (BD Biosciences Transduction Laboratories, Lexington, KY, USA).

    Techniques: Expressing, Quantitative RT-PCR, Western Blot

    Exogenous expression of miR-K12-11 partially restores HO-1 induction in ΔK12-11 mutant-infected LEC. iLEC were sequentially infected with WT or ΔK12-11 mutant KSHV for 6 days, transfected with a miR-K12-11 mimic, and then incubated for another 24 h. Samples were then harvested and prepared for various HO-1 assays. (A) qPCR analysis. For each sample, the data were normalized to the GAPDH level and the fold change was determined relative to the mock-infected sample ( n = 3). n.s., not significant; ***, P ≤ 0.0001; *, P ≤ 0.05. A one-way ANOVA was used for statistical analysis. (B) WB assay showing HO-1 protein levels. GAPDH was used as a loading control. (C) Demonstration of miR-K12-11 levels as determined by stem-loop qPCR assay. miR-K12-11 levels are normalized to the expression of the internal control miR-16 ( n = 3).

    Journal: mBio

    Article Title: Kaposi Sarcoma Herpesvirus Induces HO-1 during De Novo Infection of Endothelial Cells via Viral miRNA-Dependent and -Independent Mechanisms

    doi: 10.1128/mBio.00668-15

    Figure Lengend Snippet: Exogenous expression of miR-K12-11 partially restores HO-1 induction in ΔK12-11 mutant-infected LEC. iLEC were sequentially infected with WT or ΔK12-11 mutant KSHV for 6 days, transfected with a miR-K12-11 mimic, and then incubated for another 24 h. Samples were then harvested and prepared for various HO-1 assays. (A) qPCR analysis. For each sample, the data were normalized to the GAPDH level and the fold change was determined relative to the mock-infected sample ( n = 3). n.s., not significant; ***, P ≤ 0.0001; *, P ≤ 0.05. A one-way ANOVA was used for statistical analysis. (B) WB assay showing HO-1 protein levels. GAPDH was used as a loading control. (C) Demonstration of miR-K12-11 levels as determined by stem-loop qPCR assay. miR-K12-11 levels are normalized to the expression of the internal control miR-16 ( n = 3).

    Article Snippet: HO-1 protein levels were assessed with a mouse anti-HO-1 monoclonal antibody (MAb; BD Transduction Laboratories, Franklin Lakes, NJ) at concentrations of 1:1,000 for Western blotting (WB) and 1:200 for IFA and flow cytometry.

    Techniques: Expressing, Mutagenesis, Infection, Transfection, Incubation, Real-time Polymerase Chain Reaction, Western Blot

    KSHV miR-K12-11 is not responsible for the early peak of HO-1 induction. (A to C) Time course experiments in which iLEC were infected with KSHV and then harvested at the indicated times thereafter. (A) Stem-loop qPCR assay of miR-K12-11 in cells infected with either live or UV-inactivated virus. miR-K12-11 expression is normalized to levels of the cellular internal control miR-16 ( n = 3). (B) qPCR assay showing HO-1 mRNA levels ( n = 3) in cells infected with WT or ΔK12-11 or ΔK12-1 mutant (as a control) KSHV. (C) WB showing HO-1 protein levels in mock-infected cells (M) or cells infected with the WT or ΔK12-11 (ΔK11) or ΔK12-1 (ΔK1) mutant (control) virus. The KSHV-encoded ORF45 protein was used as a marker of infection. GAPDH served as a loading control.

    Journal: mBio

    Article Title: Kaposi Sarcoma Herpesvirus Induces HO-1 during De Novo Infection of Endothelial Cells via Viral miRNA-Dependent and -Independent Mechanisms

    doi: 10.1128/mBio.00668-15

    Figure Lengend Snippet: KSHV miR-K12-11 is not responsible for the early peak of HO-1 induction. (A to C) Time course experiments in which iLEC were infected with KSHV and then harvested at the indicated times thereafter. (A) Stem-loop qPCR assay of miR-K12-11 in cells infected with either live or UV-inactivated virus. miR-K12-11 expression is normalized to levels of the cellular internal control miR-16 ( n = 3). (B) qPCR assay showing HO-1 mRNA levels ( n = 3) in cells infected with WT or ΔK12-11 or ΔK12-1 mutant (as a control) KSHV. (C) WB showing HO-1 protein levels in mock-infected cells (M) or cells infected with the WT or ΔK12-11 (ΔK11) or ΔK12-1 (ΔK1) mutant (control) virus. The KSHV-encoded ORF45 protein was used as a marker of infection. GAPDH served as a loading control.

    Article Snippet: HO-1 protein levels were assessed with a mouse anti-HO-1 monoclonal antibody (MAb; BD Transduction Laboratories, Franklin Lakes, NJ) at concentrations of 1:1,000 for Western blotting (WB) and 1:200 for IFA and flow cytometry.

    Techniques: Infection, Real-time Polymerase Chain Reaction, Expressing, Mutagenesis, Western Blot, Marker

    The KSHV ΔK12-11 mutant induces reduced levels of HO-1 in latently infected LEC. Time course experiments show HO-1 protein and mRNA levels at 6 dpi in iLEC infected with WT or ΔK12-11 or ΔK12-1 mutant (as a control) KSHV ( n = 3). (A) qPCR assay results. n.s., not significant, **, P ≤0.01. A one-way ANOVA was used for statistical analysis. (B) WB analysis. GAPDH was used as a loading control. (C) Flow cytometric analysis of infected cells gated on GFP. (D) IFA images. Top panels: HO-1 (green), LANA (red), and DAPI (blue). Bottom panels: GFP in grayscale (same cells as those shown in top panels). Images were captured at ×60 magnification (scale bar, 30 µm).

    Journal: mBio

    Article Title: Kaposi Sarcoma Herpesvirus Induces HO-1 during De Novo Infection of Endothelial Cells via Viral miRNA-Dependent and -Independent Mechanisms

    doi: 10.1128/mBio.00668-15

    Figure Lengend Snippet: The KSHV ΔK12-11 mutant induces reduced levels of HO-1 in latently infected LEC. Time course experiments show HO-1 protein and mRNA levels at 6 dpi in iLEC infected with WT or ΔK12-11 or ΔK12-1 mutant (as a control) KSHV ( n = 3). (A) qPCR assay results. n.s., not significant, **, P ≤0.01. A one-way ANOVA was used for statistical analysis. (B) WB analysis. GAPDH was used as a loading control. (C) Flow cytometric analysis of infected cells gated on GFP. (D) IFA images. Top panels: HO-1 (green), LANA (red), and DAPI (blue). Bottom panels: GFP in grayscale (same cells as those shown in top panels). Images were captured at ×60 magnification (scale bar, 30 µm).

    Article Snippet: HO-1 protein levels were assessed with a mouse anti-HO-1 monoclonal antibody (MAb; BD Transduction Laboratories, Franklin Lakes, NJ) at concentrations of 1:1,000 for Western blotting (WB) and 1:200 for IFA and flow cytometry.

    Techniques: Mutagenesis, Infection, Real-time Polymerase Chain Reaction, Western Blot, Flow Cytometry, Immunofluorescence

    KSHV infection induces a biphasic induction of HO-1 in LEC. (A) IFA of mock-infected and KSHV-infected iLEC at 6 dpi. Top panels: LANA (red), HO-1 (green), and DAPI (blue). Bottom panels: GFP (grayscale, same cells as those shown in the top panels). The left panels correspond to mock-infected cells, and the right panels correspond to KSHV-infected cells. Images were captured at ×60 magnification (scale bar, 30 µm). (B) qPCR assay of mRNA levels for HO-1 in KSHV-infected iLEC at various times postinfection. Message levels are normalized to the GAPDH level, and fold change was calculated relative to mock-infected samples at each time point ( n = 3). (C) WB assay showing HO-1 protein levels in KSHV-infected iLEC at various times postinfection. GAPDH was used as a loading control. (D) qPCR assay of HO-1 mRNA levels in iLEC infected with either live or UV-inactivated KSHV. Cells were collected for analysis at the indicated times postinfection ( n = 3). (E) A WB showing HO-1 protein levels in iLEC infected with either live or UV-inactivated KSHV. Cells were collected at the indicated times postinfection. GAPDH was used as a loading control.

    Journal: mBio

    Article Title: Kaposi Sarcoma Herpesvirus Induces HO-1 during De Novo Infection of Endothelial Cells via Viral miRNA-Dependent and -Independent Mechanisms

    doi: 10.1128/mBio.00668-15

    Figure Lengend Snippet: KSHV infection induces a biphasic induction of HO-1 in LEC. (A) IFA of mock-infected and KSHV-infected iLEC at 6 dpi. Top panels: LANA (red), HO-1 (green), and DAPI (blue). Bottom panels: GFP (grayscale, same cells as those shown in the top panels). The left panels correspond to mock-infected cells, and the right panels correspond to KSHV-infected cells. Images were captured at ×60 magnification (scale bar, 30 µm). (B) qPCR assay of mRNA levels for HO-1 in KSHV-infected iLEC at various times postinfection. Message levels are normalized to the GAPDH level, and fold change was calculated relative to mock-infected samples at each time point ( n = 3). (C) WB assay showing HO-1 protein levels in KSHV-infected iLEC at various times postinfection. GAPDH was used as a loading control. (D) qPCR assay of HO-1 mRNA levels in iLEC infected with either live or UV-inactivated KSHV. Cells were collected for analysis at the indicated times postinfection ( n = 3). (E) A WB showing HO-1 protein levels in iLEC infected with either live or UV-inactivated KSHV. Cells were collected at the indicated times postinfection. GAPDH was used as a loading control.

    Article Snippet: HO-1 protein levels were assessed with a mouse anti-HO-1 monoclonal antibody (MAb; BD Transduction Laboratories, Franklin Lakes, NJ) at concentrations of 1:1,000 for Western blotting (WB) and 1:200 for IFA and flow cytometry.

    Techniques: Infection, Immunofluorescence, Real-time Polymerase Chain Reaction, Western Blot

    BACH1 levels and subcellular location are altered in KSHV-infected LEC. (A to C) iLEC were mock infected or infected with either the WT or ΔK12-11 mutant virus, samples were harvested at 6 dpi, and then BACH1 levels were determined. (A) qPCR analysis. The raw data for each sample were first normalized to those for GAPDH. The normalized data were then compared to those for the mock-infected sample in order to determine the fold change ( n = 6). n.s., not significant; *, P ≤0.05. Significance was determined with a Kruskal-Wallis test. (B) BACH1 WB assay. GFP expressed by the virus was used as marker of infection; GAPDH was used as a loading control. (C) IFA images. Top row, DAPI (blue) and BACH1 (green); bottom row, BACH1 (green) and LANA (red). Labels at the top of each panel correspond to the viruses used. Images were recorded at ×60 magnification; scale bar, 30 µm. (D and E) iLEC were first infected with the WT virus for 6 days and then transduced with either trans (control) or both trans and AdBACH1. Cells were harvested after a further 24-h incubation, and samples were prepared for either qPCR or WB assays ( n = 3). (D) qPCR analysis of HO-1 levels. n.s., not significant; ***, P ≤0.001; **, P ≤ 0.01. Significance was assessed with a one-way ANOVA. (E) WB showing exogenous BACH1 (FLAG) and endogenous HO-1 protein levels. GAPDH was used as a loading control.

    Journal: mBio

    Article Title: Kaposi Sarcoma Herpesvirus Induces HO-1 during De Novo Infection of Endothelial Cells via Viral miRNA-Dependent and -Independent Mechanisms

    doi: 10.1128/mBio.00668-15

    Figure Lengend Snippet: BACH1 levels and subcellular location are altered in KSHV-infected LEC. (A to C) iLEC were mock infected or infected with either the WT or ΔK12-11 mutant virus, samples were harvested at 6 dpi, and then BACH1 levels were determined. (A) qPCR analysis. The raw data for each sample were first normalized to those for GAPDH. The normalized data were then compared to those for the mock-infected sample in order to determine the fold change ( n = 6). n.s., not significant; *, P ≤0.05. Significance was determined with a Kruskal-Wallis test. (B) BACH1 WB assay. GFP expressed by the virus was used as marker of infection; GAPDH was used as a loading control. (C) IFA images. Top row, DAPI (blue) and BACH1 (green); bottom row, BACH1 (green) and LANA (red). Labels at the top of each panel correspond to the viruses used. Images were recorded at ×60 magnification; scale bar, 30 µm. (D and E) iLEC were first infected with the WT virus for 6 days and then transduced with either trans (control) or both trans and AdBACH1. Cells were harvested after a further 24-h incubation, and samples were prepared for either qPCR or WB assays ( n = 3). (D) qPCR analysis of HO-1 levels. n.s., not significant; ***, P ≤0.001; **, P ≤ 0.01. Significance was assessed with a one-way ANOVA. (E) WB showing exogenous BACH1 (FLAG) and endogenous HO-1 protein levels. GAPDH was used as a loading control.

    Article Snippet: HO-1 protein levels were assessed with a mouse anti-HO-1 monoclonal antibody (MAb; BD Transduction Laboratories, Franklin Lakes, NJ) at concentrations of 1:1,000 for Western blotting (WB) and 1:200 for IFA and flow cytometry.

    Techniques: Infection, Mutagenesis, Real-time Polymerase Chain Reaction, Western Blot, Marker, Immunofluorescence, Transduction, Incubation

    Effect of Nrf2 on GA-induced cytotoxicity. Schwann cells were treated with 500 μM GA for 24 h. (A) HO-1 mRNA expression was analyzed by real-time RT-PCR. (B) HO-1 protein expression was analyzed under a laser scanning microscope. Scale bar, 20 μm. (C) Schwann cells were transfected with control siRNA (ctrl siRNA) or Nrf2 siRNA and were treated or not treated with GA. Data are means ± S.D. of three independent experiments. *Significant difference from the value of control ( p

    Journal: Toxicology Reports

    Article Title: Glycolaldehyde induces endoplasmic reticulum stress and apoptosis in Schwann cells

    doi: 10.1016/j.toxrep.2015.10.014

    Figure Lengend Snippet: Effect of Nrf2 on GA-induced cytotoxicity. Schwann cells were treated with 500 μM GA for 24 h. (A) HO-1 mRNA expression was analyzed by real-time RT-PCR. (B) HO-1 protein expression was analyzed under a laser scanning microscope. Scale bar, 20 μm. (C) Schwann cells were transfected with control siRNA (ctrl siRNA) or Nrf2 siRNA and were treated or not treated with GA. Data are means ± S.D. of three independent experiments. *Significant difference from the value of control ( p

    Article Snippet: The primary antibodies used were anti-rabbit PERK monoclonal antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-mouse CHOP monoclonal antibody (Santa Cruz Biotechnology), and anti-rabbit HO-1 monoclonal antibody (Cell Signaling Technology), respectively.

    Techniques: Expressing, Quantitative RT-PCR, Laser-Scanning Microscopy, Transfection

    Effect of EPS on HO-1 in BAECs. BAECs were treated with EPS at the indicated concentrations for 24 h. HO-1 protein levels estimated by fluorescence microscopy studies (A) and by Western blot analysis (B). (C) HO-1 mRNA levels. Values in (B) and (C) are means±SD of three experiments. *Significant difference from the value of control ( P

    Journal: Redox Biology

    Article Title: Epalrestat increases glutathione, thioredoxin, and heme oxygenase-1 by stimulating Nrf2 pathway in endothelial cells

    doi: 10.1016/j.redox.2014.12.002

    Figure Lengend Snippet: Effect of EPS on HO-1 in BAECs. BAECs were treated with EPS at the indicated concentrations for 24 h. HO-1 protein levels estimated by fluorescence microscopy studies (A) and by Western blot analysis (B). (C) HO-1 mRNA levels. Values in (B) and (C) are means±SD of three experiments. *Significant difference from the value of control ( P

    Article Snippet: HO-1 and Trx-1 proteins were detected by reacting with phycoerythrin (PE)-conjugated anti-rabbit HO-1 monoclonal antibody (Cell Signaling Technology, Cambridge, UK) and PE-conjugated anti-mouse Trx-1 monoclonal antibody (GenWay Biotech, Inc., San Diego, CA, USA), respectively.

    Techniques: Fluorescence, Microscopy, Western Blot

    ATF4 induces expression of HO-1 in response to detachment-activated oxidative stress. ( A ) RT-PCR analysis of HO1 , GPX1 , and SOD2 in shNT and shATF4.HT1080 cells grown in suspension for 48 hours. Data are represented as mean fold change compared with attached cultures for 3 independent experiments ( n = 3, mean ± SD). * P

    Journal: The Journal of Clinical Investigation

    Article Title: ATF4-dependent induction of heme oxygenase 1 prevents anoikis and promotes metastasis

    doi: 10.1172/JCI78031

    Figure Lengend Snippet: ATF4 induces expression of HO-1 in response to detachment-activated oxidative stress. ( A ) RT-PCR analysis of HO1 , GPX1 , and SOD2 in shNT and shATF4.HT1080 cells grown in suspension for 48 hours. Data are represented as mean fold change compared with attached cultures for 3 independent experiments ( n = 3, mean ± SD). * P

    Article Snippet: Immunohistochemistry on serial tissue sections was performed with monoclonal anti-mouse HO-1 primary antibody (1:100, catalog NBP1-97507, Novus Biologicals), monoclonal anti-mouse ATF4 (1:250, clone 3E4C5, catalog 60035-1-Ig, Proteintech), and monoclonal anti-mouse LC3B (1:100, clone 5F10, catalog 0231-100, Nanotools Antibodies).

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction

    Model for involvement of HO-1 in ATF4-mediated metastasis. Detachment of tumor cells from ECM results in activation of the PERK pathway (and likely the IRE1 and ATF6 pathways), which are normally kept inactive by the endoplasmic reticulum–resident chaperone protein GRP78 (for clarity, only the PERK pathway, which is the focus of this work, is shown here). Activation of PERK mediates eIF2α phosphorylation and translational upregulation of ATF4. In turn, ATF4 activates a cytoprotective autophagy program by upregulating key autophagy genes. ATF4 also cooperates with NRF2 (directly activated by PERK) to activate the antioxidant protein HO-1 to counter increasing oxidative stress following loss of ECM. The combined action of the autophagic and antioxidant effector pathways of the PERK arm of the UPR enables tumor cells to survive longer in circulation and results in increased rates of formation of secondary metastatic sites.

    Journal: The Journal of Clinical Investigation

    Article Title: ATF4-dependent induction of heme oxygenase 1 prevents anoikis and promotes metastasis

    doi: 10.1172/JCI78031

    Figure Lengend Snippet: Model for involvement of HO-1 in ATF4-mediated metastasis. Detachment of tumor cells from ECM results in activation of the PERK pathway (and likely the IRE1 and ATF6 pathways), which are normally kept inactive by the endoplasmic reticulum–resident chaperone protein GRP78 (for clarity, only the PERK pathway, which is the focus of this work, is shown here). Activation of PERK mediates eIF2α phosphorylation and translational upregulation of ATF4. In turn, ATF4 activates a cytoprotective autophagy program by upregulating key autophagy genes. ATF4 also cooperates with NRF2 (directly activated by PERK) to activate the antioxidant protein HO-1 to counter increasing oxidative stress following loss of ECM. The combined action of the autophagic and antioxidant effector pathways of the PERK arm of the UPR enables tumor cells to survive longer in circulation and results in increased rates of formation of secondary metastatic sites.

    Article Snippet: Immunohistochemistry on serial tissue sections was performed with monoclonal anti-mouse HO-1 primary antibody (1:100, catalog NBP1-97507, Novus Biologicals), monoclonal anti-mouse ATF4 (1:250, clone 3E4C5, catalog 60035-1-Ig, Proteintech), and monoclonal anti-mouse LC3B (1:100, clone 5F10, catalog 0231-100, Nanotools Antibodies).

    Techniques: Activation Assay

    High HO-1 expression correlates with metastasis and reduced overall survival in patients. ( A ). ( B ) Probability of overall survival in glioblastoma patients with either upregulated (≥2-fold over the mean), downregulated (≤2-fold over the mean), or intermediate HO1 ), with n representing the patient number for each group. Statistical significance was calculated based on log-rank test. ( C ) Immunohistochemical analysis of serial lung sections from mice injected with shNT.HT1080 cells using antibodies against ATF4, HO-1, and LC3. Insets show areas of colocalization in the 3 detected proteins. D and E show expression of HO-1 and ATF4 from surgical specimens from primary breast tumor with known involvement of lymph node metastasis ( D ) and brain metastatic lesion from a patient with a primary sarcoma ( E ). T, tumor region; N, normal tissue. Original magnification, ×2, ×5 (insets) ( C ); ×2, ×4 (insets) ( D and E ).

    Journal: The Journal of Clinical Investigation

    Article Title: ATF4-dependent induction of heme oxygenase 1 prevents anoikis and promotes metastasis

    doi: 10.1172/JCI78031

    Figure Lengend Snippet: High HO-1 expression correlates with metastasis and reduced overall survival in patients. ( A ). ( B ) Probability of overall survival in glioblastoma patients with either upregulated (≥2-fold over the mean), downregulated (≤2-fold over the mean), or intermediate HO1 ), with n representing the patient number for each group. Statistical significance was calculated based on log-rank test. ( C ) Immunohistochemical analysis of serial lung sections from mice injected with shNT.HT1080 cells using antibodies against ATF4, HO-1, and LC3. Insets show areas of colocalization in the 3 detected proteins. D and E show expression of HO-1 and ATF4 from surgical specimens from primary breast tumor with known involvement of lymph node metastasis ( D ) and brain metastatic lesion from a patient with a primary sarcoma ( E ). T, tumor region; N, normal tissue. Original magnification, ×2, ×5 (insets) ( C ); ×2, ×4 (insets) ( D and E ).

    Article Snippet: Immunohistochemistry on serial tissue sections was performed with monoclonal anti-mouse HO-1 primary antibody (1:100, catalog NBP1-97507, Novus Biologicals), monoclonal anti-mouse ATF4 (1:250, clone 3E4C5, catalog 60035-1-Ig, Proteintech), and monoclonal anti-mouse LC3B (1:100, clone 5F10, catalog 0231-100, Nanotools Antibodies).

    Techniques: Expressing, Immunohistochemistry, Mouse Assay, Injection

    ATF4 and HO-1 promote fibrosarcoma lung colonization. ( A ) Kaplan-Meier survival analysis for mice injected by tail vein with shNT ( n = 4) or shATF4 HT1080 clones ( n = 8). *** P = 0.0002, log-rank test. ( B ) shNT ( n = 5), shATF4 ( n = 5), and ATF4 overexpressing (shATF4 + Ad-mATF4) HT1080 cells ( n = 8) were injected intravenously, and bioluminescent images were obtained. Representative images from 4 hours after injection and 8 weeks after injection are shown. ( C ) Mean radiance (photon/s/cm 2 /sr) from the chest/lung area of mice is plotted against time after injection (mean ± SEM). ( D ) shNT.HT1080 cells along with shATF4.HT1080 cells and HO-1–overexpressing cells (shATF4 + hHO-1) were injected. Mean radiance is plotted as a function of time (mean ± SEM). * P

    Journal: The Journal of Clinical Investigation

    Article Title: ATF4-dependent induction of heme oxygenase 1 prevents anoikis and promotes metastasis

    doi: 10.1172/JCI78031

    Figure Lengend Snippet: ATF4 and HO-1 promote fibrosarcoma lung colonization. ( A ) Kaplan-Meier survival analysis for mice injected by tail vein with shNT ( n = 4) or shATF4 HT1080 clones ( n = 8). *** P = 0.0002, log-rank test. ( B ) shNT ( n = 5), shATF4 ( n = 5), and ATF4 overexpressing (shATF4 + Ad-mATF4) HT1080 cells ( n = 8) were injected intravenously, and bioluminescent images were obtained. Representative images from 4 hours after injection and 8 weeks after injection are shown. ( C ) Mean radiance (photon/s/cm 2 /sr) from the chest/lung area of mice is plotted against time after injection (mean ± SEM). ( D ) shNT.HT1080 cells along with shATF4.HT1080 cells and HO-1–overexpressing cells (shATF4 + hHO-1) were injected. Mean radiance is plotted as a function of time (mean ± SEM). * P

    Article Snippet: Immunohistochemistry on serial tissue sections was performed with monoclonal anti-mouse HO-1 primary antibody (1:100, catalog NBP1-97507, Novus Biologicals), monoclonal anti-mouse ATF4 (1:250, clone 3E4C5, catalog 60035-1-Ig, Proteintech), and monoclonal anti-mouse LC3B (1:100, clone 5F10, catalog 0231-100, Nanotools Antibodies).

    Techniques: Mouse Assay, Injection, Clone Assay

    Area under the curve (AUC) over 48 h showing changes from individual baseline levels (ΔAUC48) for haem oxygenase (HO-1) mRNA and protein levels. Box and whisker plots of the ΔAUC48 for the HO-1 mRNA (A) and of the ODs of the protein levels

    Journal: British Journal of Pharmacology

    Article Title: Haem arginate infusion stimulates haem oxygenase-1 expression in healthy subjects

    doi: 10.1111/j.1476-5381.2010.00990.x

    Figure Lengend Snippet: Area under the curve (AUC) over 48 h showing changes from individual baseline levels (ΔAUC48) for haem oxygenase (HO-1) mRNA and protein levels. Box and whisker plots of the ΔAUC48 for the HO-1 mRNA (A) and of the ODs of the protein levels

    Article Snippet: The membranes were blocked in 5% milk in tris-buffered saline containing 0.05 % Tween 20 (TBST; Bio-Rad, Hercules, CA, USA) for 1 h, washed three times for 5 min with TBST, incubated with the primary mouse monoclonal anti-human HO-1 antibody (Clone OSA110, Assay Designs, Ann Arbor, MI, USA) overnight at 4°C, and washed and incubated with the second antibody (anti-mouse Ig horseradish peroxidase linked, Amersham; 1:20.000) for 1 h. The membranes were incubated for 5 min in the substrate solution (ECL Plus Western Blotting Detection System; Amersham Pharmacia Biotech, Piscataway, NJ, USA) and imaged using clear X-ray films (Thermo Scientific).

    Techniques: Whisker Assay

    Haem arginate (HA) infusion induces haem oxygenase (HO-1) expression in peripheral blood mononuclear cell in vivo . HO-1 mRNA (A) and protein (C) expression after treatment with different doses of HA; bars show SD. (B) Representative HO-1 Western blots

    Journal: British Journal of Pharmacology

    Article Title: Haem arginate infusion stimulates haem oxygenase-1 expression in healthy subjects

    doi: 10.1111/j.1476-5381.2010.00990.x

    Figure Lengend Snippet: Haem arginate (HA) infusion induces haem oxygenase (HO-1) expression in peripheral blood mononuclear cell in vivo . HO-1 mRNA (A) and protein (C) expression after treatment with different doses of HA; bars show SD. (B) Representative HO-1 Western blots

    Article Snippet: The membranes were blocked in 5% milk in tris-buffered saline containing 0.05 % Tween 20 (TBST; Bio-Rad, Hercules, CA, USA) for 1 h, washed three times for 5 min with TBST, incubated with the primary mouse monoclonal anti-human HO-1 antibody (Clone OSA110, Assay Designs, Ann Arbor, MI, USA) overnight at 4°C, and washed and incubated with the second antibody (anti-mouse Ig horseradish peroxidase linked, Amersham; 1:20.000) for 1 h. The membranes were incubated for 5 min in the substrate solution (ECL Plus Western Blotting Detection System; Amersham Pharmacia Biotech, Piscataway, NJ, USA) and imaged using clear X-ray films (Thermo Scientific).

    Techniques: Expressing, In Vivo, Western Blot