anti-ho-1 Search Results


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  • 93
    Thermo Fisher anti ho 1
    PP upregulates the expression of <t>HO-1</t> in the lungs of mice. (A) Expression levels of HO-1 were examined by western blot analysis. (B) Quantitative analysis of HO-1 was performed by densitometric analysis. ** P
    Anti Ho 1, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Millipore anti ho 1
    Glutamate dehydrogenase and heme oxygenase-1. A: Glutamate dehydrogenase (GLDH) levels after 120 min of reperfusion; B: Heme oxygenase-1 <t>(HO-1)</t> protein levels in liver after 120 min of normothermic reperfusion. Representative Western blotting at (top) and densitometric analysis at (bottom). a P
    Anti Ho 1, supplied by Millipore, used in various techniques. Bioz Stars score: 94/100, based on 115 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Abcam anti ho 1
    Double immunofluorescence staining assay for the co-expression in glia cells. A. Immunofluorescence merge images for co-expressing of <t>HO-1,</t> Nrf2, NF-κB and TNF-α with the CD11b in glia cells. B. Statistical analysis for the co-expression
    Anti Ho 1, supplied by Abcam, used in various techniques. Bioz Stars score: 95/100, based on 416 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Santa Cruz Biotechnology anti ho 1
    Effects of HMGB1 on the expressions of VAP-1, 8-OHdG, and <t>HO-1</t> in human retinal microvascular endothelial cells. Immunofluorescence microscopy showed that HMGB1 treatment induced membranous VAP-1 translocation ( A ), the upregulation of cytoplasmic and nuclear immunoreactivities for 8-OHdG ( B ), and cytoplasmic immunoreactivity for HO-1 ( C ; green). Nuclei were counterstained with DAPI (blue).
    Anti Ho 1, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 95/100, based on 438 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti ho 1/product/Santa Cruz Biotechnology
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    92
    Stressgen Biotechnologies anti ho 1
    Effects of clofarabine and resveratrol on Akt and Erk1/2 phosphorylation. (A) MSTO-211H cells were treated with clofarabine (40 nM) and resveratrol (15 μM) for the indicated times. (B) Cells were pretreated with or without Ly294002 (20 μM), PD98059 (50 nM), and NAC (5 mM) for 1 h prior to combined treatment for 24 h. Cell lysates were analyzed by immunoblotting using anti-p-Akt, anti-p-Erk, anti-Nrf2, and <t>anti-HO-1</t> antibodies. (C) Cells were treated with or without NAC (5 mM) for 1 h prior to the combination treatment for 24 h. The percentage of viable cells was then determined by the trypan blue exclusion assay. *P < 0.05 compared with respective controls.
    Anti Ho 1, supplied by Stressgen Biotechnologies, used in various techniques. Bioz Stars score: 92/100, based on 76 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    R&D Systems anti ho 1
    Activation of H-Ras induces <t>HO-1</t> promoter activity: A and B , Caki-1 and 786-0 cells were co-transfected with the HO-1 promoter-luciferase construct (0.5 μg) and either different amounts (0.5 and 1.0 μg) of active H-Ras overexpression plasmid
    Anti Ho 1, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    GeneTex anti ho 1
    Effect of lucidone on <t>HO-1</t> expression. (A to C) Concentration-dependent induction of HO-1 promoter activity (A), RNA transcription (B), and protein synthesis by lucidone (C). For the promoter activity assay, Ava5 cells were transiently transfected with
    Anti Ho 1, supplied by GeneTex, used in various techniques. Bioz Stars score: 90/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Enzo Biochem anti ho 1
    Effect of SIN on oxidative stress. After treatment with 0.25, 0.5, or 1 mM for 24 h, the level of MDA ( A ) and activity of SOD ( B ) and CAT ( C ) was detected; Protein ( D ) and mRNA ( E ) levels of Nrf2, <t>HO-1,</t> and NQO-1 were detected using Western blotting and qRT-PCR, respectively. Controls: cells without any treatment; BK: 1 μg/ml BK treatment group; BK + SIN 0.25: 1 μg/ml BK + 0.25 nM SIN treatment group; BK + SIN 0.5: 1 μg/ml BK + 0.5 nM SIN treatment group; BK + SIN 1: 1 μg/ml BK + 1 nM SIN treatment group; * p
    Anti Ho 1, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 93/100, based on 138 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Epitomics anti ho 1
    Flaccidoside II downregulates the expression and activation of antiapoptotic <t>HO-1,</t> which is involved in the apoptotic facilitation of flaccidoside II. Notes: ( A ) The ST88-14 and S462 cells were treated with either 40.0 μmol L −1 flaccidoside II or DMSO for 48 hours, and the protein expression level of HO-1 was measured by Western blotting, which was normalized to GAPDH. ( B ) The HO activity was measured in ST88-14 and S462 cells at 48 hours after treatment of these cells with 40.0 μmol L −1 flaccidoside II or DMSO. ( C ) The ST88-14 and S462 cells were treated with 40.0 μmol L −1 flaccidoside II with the transfection of either vector plasmid as a negative control or full-length HO-1 plasmid. The expression of HO-1, C-caspase 3, total PARP, and cleaved PARP were determined by Western blotting, and GAPDH was used as loading control ( C ). ( D ) Quantitative analysis of the apoptosis markers cleaved caspase 3 and cleaved PARP in ( C ). ( E ) The apoptotic rates of ST88-14 and S462 cells were determined by Annexin V-PI staining and flow cytometry for quantification. Each bar represents the mean ± SD of four independent experiments. *** P
    Anti Ho 1, supplied by Epitomics, used in various techniques. Bioz Stars score: 92/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Becton Dickinson anti ho 1
    KMUP-1 treatment elevated the expression of <t>HO-1</t> protein. ( A ) H9c2 cells were preincubated with ZnPP at 10 μM for 1 h, followed by co-treatment with KMUP-1 or ( B ) combination with 100 nM of ET-1. Western blot analysis was conducted to measure the induction of HO-1 protein; ( C ) The expression of calcineurin A was analyzed. # p
    Anti Ho 1, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 91/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Abnova anti ho 1
    a Quantification of densitometric analysis of immunohistochemistry. Data are presented as median (IQR). Representative pictures ×10. b CSE YGP, c CSE FBM sham, d CSE FBM sepsis, e <t>HO-1</t> YGP, f HO-1 FBM sham, and ( g ) HO-1 FBM sepsis. b – g show a cross-section of the vessel wall with the luminal side to the right . h eNOS YGP, i eNOS FBM sham, and ( j ) eNOS FBM sepsis. h – j show a cross-section of the vessel wall with the luminal side to the upper left corner
    Anti Ho 1, supplied by Abnova, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    StressMarq anti ho 1
    DMF  increases expression of antioxidant enzymes in  RAW  264.7 cells. Gene expression for  NQO 1  ( A ),  HO ‐1  ( B ) and  GCS  ( C ) are shown. * P 
    Anti Ho 1, supplied by StressMarq, used in various techniques. Bioz Stars score: 91/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Assay Designs Inc anti ho 1
    Intracellular heme oxygenase 1 <t>(HO-1)</t> in primary human tracheo-bronchial epithelial cells 24 h after treatment with sulfur mustard (SM) vapor. HO-1 levels in the medium were measured by western blotting, and normalized to β-actin levels in the same blots following stripping and re-probing. Incubator control (inc ctl): cells maintained in the CO 2 incubator during exposure of the remaining cultures in a glove box. 0: Cells exposed to the vapor from 10 μl of ethanol. SM concentrations are the estimated maximal concentration possible in the vapor phase as the result of the amount present in 10 μl of the dilution applied to the filter.
    Anti Ho 1, supplied by Assay Designs Inc, used in various techniques. Bioz Stars score: 92/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Sangon Biotech anti ho 1
    SFN pretreatment and cadmium-induced Nrf2/ARE (antioxidant response element) signaling in mouse testes. ( A ) Nuclear factor-erythroid 2-related factor 2 (Nrf2); ( B ) NAD(P)H:quinone oxidoreductase 1 (NQO1); ( C ) hemeoxygenase-1 <t>(HO-1);</t> ( D ) γ-glutamyl cysteine synthetase (γ-GCS); and ( E ) glutathione peroxidase (GSH-Px). Values represent mean ± SEM in each group of 10 mice; * p
    Anti Ho 1, supplied by Sangon Biotech, used in various techniques. Bioz Stars score: 92/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Enzo Biochem polyclonal anti ho 1
    SFN pretreatment and cadmium-induced Nrf2/ARE (antioxidant response element) signaling in mouse testes. ( A ) Nuclear factor-erythroid 2-related factor 2 (Nrf2); ( B ) NAD(P)H:quinone oxidoreductase 1 (NQO1); ( C ) hemeoxygenase-1 <t>(HO-1);</t> ( D ) γ-glutamyl cysteine synthetase (γ-GCS); and ( E ) glutathione peroxidase (GSH-Px). Values represent mean ± SEM in each group of 10 mice; * p
    Polyclonal Anti Ho 1, supplied by Enzo Biochem, used in various techniques. Bioz Stars score: 88/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Proteintech anti ho 1 antibodies
    DMF suppresses LPS-induced reactive MG. Primary MG were stimulated by LPS (100 ng/ml) in the absence or presence of DMF for various time periods. a Microglial surface markers, CD86, CD80, and CD40, were examined at 18 h after stimulation using FACS analysis. Representative flow plots from five independent experiments are shown. ISO, the isotype control antibody. MED, cell culture medium. b Morphological changes of MG at 18 h after stimulation were examined using Iba1 immunostaining. Arrows indicate activated MG with increased expression of Iba1 and enlarged cell bodies. Cell nuclei were stained blue with Hoechst 33342. Scale bar, 20 μm. c The expressions of DMF-induced Nrf2 target genes were examined at 3 h after stimulation. The mRNA levels of <t>hmox1</t> , nqo1 , gclc , and gclm were quantified using qPCR. Data presented are from 3 to 5 independent experiments. * p
    Anti Ho 1 Antibodies, supplied by Proteintech, 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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    Image Search Results


    PP upregulates the expression of HO-1 in the lungs of mice. (A) Expression levels of HO-1 were examined by western blot analysis. (B) Quantitative analysis of HO-1 was performed by densitometric analysis. ** P

    Journal: International Journal of Molecular Medicine

    Article Title: Physalis peruviana L. inhibits ovalbumin-induced airway inflammation by attenuating the activation of NF-κB and inflammatory molecules

    doi: 10.3892/ijmm.2019.4110

    Figure Lengend Snippet: PP upregulates the expression of HO-1 in the lungs of mice. (A) Expression levels of HO-1 were examined by western blot analysis. (B) Quantitative analysis of HO-1 was performed by densitometric analysis. ** P

    Article Snippet: The primary antibodies and dilution rates were as follows: Anti-phosphorylated (p)-p38 (cat. no. 9211), anti-p-extracellular signal-regulated kinase (ERK; cat. no. 4370), anti-p-NF-κB p65 (cat. no. 3033) and anti-β-actin (cat. no. 4967) at 1:1,000 dilutions (Cell Signaling Technology, Inc., Danvers, MA, USA), anti-MCP-1 (cat. no. sc-28879), anti-p38 (cat. no. sc-7149), anti-ERK (cat. no. sc-154), anti-p-c-Jun N-terminal kinase (JNK; cat. no. sc-6254), anti-JNK (cat. no. sc-474), anti-NF-κB p65 (cat. no. sc-372), anti-inhibitor of NF-κB (IκB-α; cat. no. sc-1643) and anti-KEN-5 (cat. no. sc-59373) at 1:1,000 dilutions (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), and anti-HO-1 (cat. no. PA5-27338; Thermo Fisher Scientific, Inc.) at 1:1,000 dilution.

    Techniques: Expressing, Mouse Assay, Western Blot

    Glutamate dehydrogenase and heme oxygenase-1. A: Glutamate dehydrogenase (GLDH) levels after 120 min of reperfusion; B: Heme oxygenase-1 (HO-1) protein levels in liver after 120 min of normothermic reperfusion. Representative Western blotting at (top) and densitometric analysis at (bottom). a P

    Journal: World Journal of Gastroenterology : WJG

    Article Title: Hypoxia inducible factor-1? accumulation in steatotic liver preservation: Role of nitric oxide

    doi: 10.3748/wjg.v16.i28.3499

    Figure Lengend Snippet: Glutamate dehydrogenase and heme oxygenase-1. A: Glutamate dehydrogenase (GLDH) levels after 120 min of reperfusion; B: Heme oxygenase-1 (HO-1) protein levels in liver after 120 min of normothermic reperfusion. Representative Western blotting at (top) and densitometric analysis at (bottom). a P

    Article Snippet: Membranes were immunoblotted with antibodies against anti-eNOS (transduction laboratories, Lexington, KY, USA), against anti-HO-1 (Sigma Chemical, St Louis, MO, USA) and β-actin (Sigma Chemical, St. Louis, MO, USA).

    Techniques: Western Blot

    Double immunofluorescence staining assay for the co-expression in glia cells. A. Immunofluorescence merge images for co-expressing of HO-1, Nrf2, NF-κB and TNF-α with the CD11b in glia cells. B. Statistical analysis for the co-expression

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Heme oxygenase 1 plays role of neuron-protection by regulating Nrf2-ARE signaling post intracerebral hemorrhage

    doi:

    Figure Lengend Snippet: Double immunofluorescence staining assay for the co-expression in glia cells. A. Immunofluorescence merge images for co-expressing of HO-1, Nrf2, NF-κB and TNF-α with the CD11b in glia cells. B. Statistical analysis for the co-expression

    Article Snippet: The SDS page isolated proteins were incubated overnight at 4°C with the anti-Nrf2 (1:000, Abcam, U. K.), anti-HO-1 (1:1000, Abcam, UK), anti-NF-κB (1:1000, Cell Signaling Technology, U. S.), anti-TNF-α (1:3000, Santa Cruz, USA) and anti-β-actin (1:1000, Sigma, USA).

    Techniques: Double Immunofluorescence Staining, Expressing, Immunofluorescence

    mRNA expression of HO-1, Nrf2, NF-κB and TNF-α in ICH and ZPP-IX group at different time. A. mRNA expression of HO-1 gene. B. mRNA expression of Nrf2 gene. C. mRNA expression of NF-κB gene. D. mRNA expression of TNF-α gene.

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Heme oxygenase 1 plays role of neuron-protection by regulating Nrf2-ARE signaling post intracerebral hemorrhage

    doi:

    Figure Lengend Snippet: mRNA expression of HO-1, Nrf2, NF-κB and TNF-α in ICH and ZPP-IX group at different time. A. mRNA expression of HO-1 gene. B. mRNA expression of Nrf2 gene. C. mRNA expression of NF-κB gene. D. mRNA expression of TNF-α gene.

    Article Snippet: The SDS page isolated proteins were incubated overnight at 4°C with the anti-Nrf2 (1:000, Abcam, U. K.), anti-HO-1 (1:1000, Abcam, UK), anti-NF-κB (1:1000, Cell Signaling Technology, U. S.), anti-TNF-α (1:3000, Santa Cruz, USA) and anti-β-actin (1:1000, Sigma, USA).

    Techniques: Expressing

    HO-1 expression in ICH and ZPP-IX group. A. Western blot assay for the HO-1 expression. B. Statistical analysis for the HO-1 protein. * P

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Heme oxygenase 1 plays role of neuron-protection by regulating Nrf2-ARE signaling post intracerebral hemorrhage

    doi:

    Figure Lengend Snippet: HO-1 expression in ICH and ZPP-IX group. A. Western blot assay for the HO-1 expression. B. Statistical analysis for the HO-1 protein. * P

    Article Snippet: The SDS page isolated proteins were incubated overnight at 4°C with the anti-Nrf2 (1:000, Abcam, U. K.), anti-HO-1 (1:1000, Abcam, UK), anti-NF-κB (1:1000, Cell Signaling Technology, U. S.), anti-TNF-α (1:3000, Santa Cruz, USA) and anti-β-actin (1:1000, Sigma, USA).

    Techniques: Expressing, Western Blot

    The gene expression of cytokines on Nrf2 pathway in livers from Sprague Dawley rats after SCU treatmentby quantitative RT-PCR.Quantitative values were obtained from the threshold cycle value ( ΔΔ Ct). ( A–C ) The quantitative values of HO-1, NQO1, Nrf2 gene expression in different groups.All data were represented as means±standard deviation (n=3).Statistical analysis of these date (* P

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

    Article Title: Scutellarin Prevents Nonalcoholic Fatty Liver Disease (NAFLD) and Hyperlipidemia via PI3K/AKT-Dependent Activation of Nuclear Factor (Erythroid-Derived 2)-Like 2 (Nrf2) in Rats

    doi: 10.12659/MSM.907530

    Figure Lengend Snippet: The gene expression of cytokines on Nrf2 pathway in livers from Sprague Dawley rats after SCU treatmentby quantitative RT-PCR.Quantitative values were obtained from the threshold cycle value ( ΔΔ Ct). ( A–C ) The quantitative values of HO-1, NQO1, Nrf2 gene expression in different groups.All data were represented as means±standard deviation (n=3).Statistical analysis of these date (* P

    Article Snippet: Anti-PI3K and anti-AKT (phospho Ser473) were acquired from Immunoway Biotechnology Company (USA), anti-Nrf2 antibody (AP52270) was purchased from Abgent (USA), NQO1 (ab28947) and anti-HO-1 antibody (ab69544) were acquired from ABcam Co. LTD (Shanghai).

    Techniques: Expressing, Quantitative RT-PCR, Standard Deviation

    The evaluation on Nrf2-mediated antioxidant defense system of livers from Sprague Dawley rats after SCU treatment by western blotting. The levels of AKT/Nrf2 pathway proteins were evaluated by western blotting. β-actin was used as the internal control, respectively. ( A ) HO-1, ( B ) NQO1, ( C ) Nrf2, ( D ) p-AKT, and ( E ) PI3K proteins in different groups. All data were represented as means ± standard deviation (n=6).Statistical analysis of these date. (* P

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

    Article Title: Scutellarin Prevents Nonalcoholic Fatty Liver Disease (NAFLD) and Hyperlipidemia via PI3K/AKT-Dependent Activation of Nuclear Factor (Erythroid-Derived 2)-Like 2 (Nrf2) in Rats

    doi: 10.12659/MSM.907530

    Figure Lengend Snippet: The evaluation on Nrf2-mediated antioxidant defense system of livers from Sprague Dawley rats after SCU treatment by western blotting. The levels of AKT/Nrf2 pathway proteins were evaluated by western blotting. β-actin was used as the internal control, respectively. ( A ) HO-1, ( B ) NQO1, ( C ) Nrf2, ( D ) p-AKT, and ( E ) PI3K proteins in different groups. All data were represented as means ± standard deviation (n=6).Statistical analysis of these date. (* P

    Article Snippet: Anti-PI3K and anti-AKT (phospho Ser473) were acquired from Immunoway Biotechnology Company (USA), anti-Nrf2 antibody (AP52270) was purchased from Abgent (USA), NQO1 (ab28947) and anti-HO-1 antibody (ab69544) were acquired from ABcam Co. LTD (Shanghai).

    Techniques: Western Blot, Standard Deviation

    Suppressive and proliferative function of Tregs in healthy controls and vitiligo patients after HO‐1 induction by Hemin. (A and B) Representative FACS chart for LAP expression on Tregs before and after Hemin stimulation, and the relevant statistical comparisons, as indicated. (C and D) Representative FACS chart for IL‐10 expression on Tregs before and after Hemin stimulation, and the relevant statistical comparisons. (E and F) Representative FACS chart for CTLA‐4 expression on Tregs before and after Hemin stimulation and the relevant statistical comparisons. (G and H) Representative FACS chart for Treg suppressive ability analysis before and after Hemin treatment in healthy controls and vitiligo patients, and the relevant statistical comparisons. (I and J) Representative FACS chart for Treg proliferation ability analysis before and after Hemin treatment in healthy controls and vitiligo patients, and the relevant statistical comparisons. Values are presented as the mean ± SD, * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: HO‐1 regulates the function of Treg: Association with the immune intolerance in vitiligo, et al. HO‐1 regulates the function of Treg: Association with the immune intolerance in vitiligo

    doi: 10.1111/jcmm.13723

    Figure Lengend Snippet: Suppressive and proliferative function of Tregs in healthy controls and vitiligo patients after HO‐1 induction by Hemin. (A and B) Representative FACS chart for LAP expression on Tregs before and after Hemin stimulation, and the relevant statistical comparisons, as indicated. (C and D) Representative FACS chart for IL‐10 expression on Tregs before and after Hemin stimulation, and the relevant statistical comparisons. (E and F) Representative FACS chart for CTLA‐4 expression on Tregs before and after Hemin stimulation and the relevant statistical comparisons. (G and H) Representative FACS chart for Treg suppressive ability analysis before and after Hemin treatment in healthy controls and vitiligo patients, and the relevant statistical comparisons. (I and J) Representative FACS chart for Treg proliferation ability analysis before and after Hemin treatment in healthy controls and vitiligo patients, and the relevant statistical comparisons. Values are presented as the mean ± SD, * P

    Article Snippet: The following mAb were used: anti‐Foxp3‐APC (eBioscience), anti‐HO‐1‐PE (Abcam), anti‐IL‐10‐PE (eBioscience), anti‐LAP‐PerCP (eBioscience) and anti‐CTLA‐4‐PE (Biolegend).

    Techniques: FACS, Expressing

    HO‐1 and its reactive products level in healthy controls and vitiligo patients. (A and B) Representative FACS plots and relevant statistical comparisons of HO‐1 expression in Tregs of vitiligo patients and healthy controls (n = 20). (C) Elisa assay for serum level of HO‐1 (n = 20). (D) Elisa assay for serum level of CoHb (n = 20). (E) Elisa assay for serum level of TBIL (n = 20). (F) Elisa assay for serum level of Fe 2+ (n = 20). Values are presented as the mean ± SD, ** P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: HO‐1 regulates the function of Treg: Association with the immune intolerance in vitiligo, et al. HO‐1 regulates the function of Treg: Association with the immune intolerance in vitiligo

    doi: 10.1111/jcmm.13723

    Figure Lengend Snippet: HO‐1 and its reactive products level in healthy controls and vitiligo patients. (A and B) Representative FACS plots and relevant statistical comparisons of HO‐1 expression in Tregs of vitiligo patients and healthy controls (n = 20). (C) Elisa assay for serum level of HO‐1 (n = 20). (D) Elisa assay for serum level of CoHb (n = 20). (E) Elisa assay for serum level of TBIL (n = 20). (F) Elisa assay for serum level of Fe 2+ (n = 20). Values are presented as the mean ± SD, ** P

    Article Snippet: The following mAb were used: anti‐Foxp3‐APC (eBioscience), anti‐HO‐1‐PE (Abcam), anti‐IL‐10‐PE (eBioscience), anti‐LAP‐PerCP (eBioscience) and anti‐CTLA‐4‐PE (Biolegend).

    Techniques: FACS, Expressing, Enzyme-linked Immunosorbent Assay

    Effects of HMGB1 on the expressions of VAP-1, 8-OHdG, and HO-1 in human retinal microvascular endothelial cells. Immunofluorescence microscopy showed that HMGB1 treatment induced membranous VAP-1 translocation ( A ), the upregulation of cytoplasmic and nuclear immunoreactivities for 8-OHdG ( B ), and cytoplasmic immunoreactivity for HO-1 ( C ; green). Nuclei were counterstained with DAPI (blue).

    Journal: Molecular Vision

    Article Title: Association of HMGB1 with oxidative stress markers and regulators in PDR

    doi:

    Figure Lengend Snippet: Effects of HMGB1 on the expressions of VAP-1, 8-OHdG, and HO-1 in human retinal microvascular endothelial cells. Immunofluorescence microscopy showed that HMGB1 treatment induced membranous VAP-1 translocation ( A ), the upregulation of cytoplasmic and nuclear immunoreactivities for 8-OHdG ( B ), and cytoplasmic immunoreactivity for HO-1 ( C ; green). Nuclei were counterstained with DAPI (blue).

    Article Snippet: They were left untreated or were treated with 100 ng/ml of cytokine-HMGB1, lipopolysaccharide free (Cat. No. REHM120, IBL International Corporation) for 24 h and were fixed with ice-cold acetone or 4% paraformaldehyde followed by blocking with 10% goat serum for 1 h. Without being washed, the cells were incubated with anti-HO-1 (1:100, sc-10789, Santa Cruz Biotechnology, Inc.), anti-VAP-1 (1:100, ab196739, Abcam), or anti-8OHdG (1:50, ab62623, Abcam) primary antibodies overnight at 4 °C.

    Techniques: Immunofluorescence, Microscopy, Translocation Assay

    Association of Brg1 overexpression with activated antioxidant and decreased inflammatory factors. (a–c) the protein expression of HO-1 and NQO1 in the CMV-Brg1 and WT mice after HIR. (d–h) the concentrations of glutamate-cysteine ligase catalytic subunit (GCLC), glutathione S-transferase alpha 1 (GST α 1), superoxide dismutase (SOD), interleukin-6 (IL-6), and tumor necrosis factor- α (TNF- α ) in the CMV-Brg1 and WT mice after HIR. ∗ P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Overexpression of Brg1 Alleviates Hepatic Ischemia/Reperfusion-Induced Acute Lung Injury through Antioxidative Stress Effects

    doi: 10.1155/2017/8787392

    Figure Lengend Snippet: Association of Brg1 overexpression with activated antioxidant and decreased inflammatory factors. (a–c) the protein expression of HO-1 and NQO1 in the CMV-Brg1 and WT mice after HIR. (d–h) the concentrations of glutamate-cysteine ligase catalytic subunit (GCLC), glutathione S-transferase alpha 1 (GST α 1), superoxide dismutase (SOD), interleukin-6 (IL-6), and tumor necrosis factor- α (TNF- α ) in the CMV-Brg1 and WT mice after HIR. ∗ P

    Article Snippet: Anti-Nrf2 antibody (1 : 100, AP52269, Abgent), anti-Brg1 antibody (1 : 100; ab110641, Abcam), anti-NQO1 antibody (1 : 100; ab28947, Abcam), anti-HO-1 antibody (1 : 100; sc-10,789, Santa Cruz), and secondary antibody (1 : 2000; Millipore) were used to detect protein expression.

    Techniques: Over Expression, Expressing, Mouse Assay

    Effects of clofarabine and resveratrol on Akt and Erk1/2 phosphorylation. (A) MSTO-211H cells were treated with clofarabine (40 nM) and resveratrol (15 μM) for the indicated times. (B) Cells were pretreated with or without Ly294002 (20 μM), PD98059 (50 nM), and NAC (5 mM) for 1 h prior to combined treatment for 24 h. Cell lysates were analyzed by immunoblotting using anti-p-Akt, anti-p-Erk, anti-Nrf2, and anti-HO-1 antibodies. (C) Cells were treated with or without NAC (5 mM) for 1 h prior to the combination treatment for 24 h. The percentage of viable cells was then determined by the trypan blue exclusion assay. *P < 0.05 compared with respective controls.

    Journal: BMB Reports

    Article Title: Synergistic inhibition of mesothelioma cell growth by the combination of clofarabine and resveratrol involves Nrf2 downregulation

    doi: 10.5483/BMBRep.2012.45.11.111

    Figure Lengend Snippet: Effects of clofarabine and resveratrol on Akt and Erk1/2 phosphorylation. (A) MSTO-211H cells were treated with clofarabine (40 nM) and resveratrol (15 μM) for the indicated times. (B) Cells were pretreated with or without Ly294002 (20 μM), PD98059 (50 nM), and NAC (5 mM) for 1 h prior to combined treatment for 24 h. Cell lysates were analyzed by immunoblotting using anti-p-Akt, anti-p-Erk, anti-Nrf2, and anti-HO-1 antibodies. (C) Cells were treated with or without NAC (5 mM) for 1 h prior to the combination treatment for 24 h. The percentage of viable cells was then determined by the trypan blue exclusion assay. *P < 0.05 compared with respective controls.

    Article Snippet: The membranes were incubated for 2 h at room temperature with a 1:500 dilution of anti-Nrf2 (Santa Cruz Biotechnology), anti-HO-1 (Stressgen Biotechnologies), anti-p-Akt, and anti-p-Erk antibodies (Cell Signaling Technologies).

    Techniques: Trypan Blue Exclusion Assay

    HO-1 overexpressing mice are protected against pulmonary inflammation induced by hypoxia. ( a ) Immunostaining for neutrophils and macrophages. Frozen lung sections from Tg or non-Tg control mice that had been exposed to hypoxia for 48 h were stained with anti-Ly-6G antibody for neutrophils and anti-Mac-3 antibody for macrophages (brown staining). Original magnification: ×100. ( b ) The number of neutrophils and macrophages in BAL fluid after hypoxia. Differential cell counts for neutrophils (Neut, Upper ) and macrophages (Mac, Lower ) were performed in the BAL fluid of Tg (closed bars) and non-Tg control mice (open bars) after 48 h of hypoxia. Data are expressed as mean ± SEM ( n = 4). *, P

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

    Article Title: Targeted expression of heme oxygenase-1 prevents the pulmonary inflammatory and vascular responses to hypoxia

    doi: 10.1073/pnas.161272598

    Figure Lengend Snippet: HO-1 overexpressing mice are protected against pulmonary inflammation induced by hypoxia. ( a ) Immunostaining for neutrophils and macrophages. Frozen lung sections from Tg or non-Tg control mice that had been exposed to hypoxia for 48 h were stained with anti-Ly-6G antibody for neutrophils and anti-Mac-3 antibody for macrophages (brown staining). Original magnification: ×100. ( b ) The number of neutrophils and macrophages in BAL fluid after hypoxia. Differential cell counts for neutrophils (Neut, Upper ) and macrophages (Mac, Lower ) were performed in the BAL fluid of Tg (closed bars) and non-Tg control mice (open bars) after 48 h of hypoxia. Data are expressed as mean ± SEM ( n = 4). *, P

    Article Snippet: Sections were treated with 0.3% hydrogen peroxide in methanol for 20 min, preincubated with 5% goat serum, and treated with the anti-Ly-6G antibody (1:500, PharMingen) for neutrophils, anti-Mac-3 antibody for macrophages (1: 500, PharMingen), or anti-HO-1 antibody (1:200, SPA 896, StressGen Biotechnologies) for 1 h at 37°C.

    Techniques: Mouse Assay, Immunostaining, Staining

    Immunostaining for HO-1 in the lung of Tg mice. Paraffin-embedded sections of the lungs were analyzed for HO-1 expression by immunohistochemistry. ( a ) High levels of HO-1 expression are evident in the lung of Tg mice (brown, arrow). ( b ) High power photograph of the rectangle area indicated in a . ( c ) The section adjacent to that in a incubated with preimmune rabbit serum. ( d ) Absence of detectable HO-1 immunoreactivity in the lung of non-Tg mice. Original magnification: ×100 for a , c , and d and ×400 for b . Staining was performed in lung sections of four Tg and four non-Tg control mice.

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

    Article Title: Targeted expression of heme oxygenase-1 prevents the pulmonary inflammatory and vascular responses to hypoxia

    doi: 10.1073/pnas.161272598

    Figure Lengend Snippet: Immunostaining for HO-1 in the lung of Tg mice. Paraffin-embedded sections of the lungs were analyzed for HO-1 expression by immunohistochemistry. ( a ) High levels of HO-1 expression are evident in the lung of Tg mice (brown, arrow). ( b ) High power photograph of the rectangle area indicated in a . ( c ) The section adjacent to that in a incubated with preimmune rabbit serum. ( d ) Absence of detectable HO-1 immunoreactivity in the lung of non-Tg mice. Original magnification: ×100 for a , c , and d and ×400 for b . Staining was performed in lung sections of four Tg and four non-Tg control mice.

    Article Snippet: Sections were treated with 0.3% hydrogen peroxide in methanol for 20 min, preincubated with 5% goat serum, and treated with the anti-Ly-6G antibody (1:500, PharMingen) for neutrophils, anti-Mac-3 antibody for macrophages (1: 500, PharMingen), or anti-HO-1 antibody (1:200, SPA 896, StressGen Biotechnologies) for 1 h at 37°C.

    Techniques: Immunostaining, Mouse Assay, Expressing, Immunohistochemistry, Incubation, Staining

    Time course of hypoxic right ventricular hypertrophy: protection by HO-1 overexpression. ( a ) Non-Tg mice were exposed to hypoxia (8–10% O 2 ) for 18 h, 5 days (d), or 2–3 weeks (w). Development of RVH after hypoxic exposure was determined by the RV/(LV + S) ratio as described in Materials and Methods . *, P

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

    Article Title: Targeted expression of heme oxygenase-1 prevents the pulmonary inflammatory and vascular responses to hypoxia

    doi: 10.1073/pnas.161272598

    Figure Lengend Snippet: Time course of hypoxic right ventricular hypertrophy: protection by HO-1 overexpression. ( a ) Non-Tg mice were exposed to hypoxia (8–10% O 2 ) for 18 h, 5 days (d), or 2–3 weeks (w). Development of RVH after hypoxic exposure was determined by the RV/(LV + S) ratio as described in Materials and Methods . *, P

    Article Snippet: Sections were treated with 0.3% hydrogen peroxide in methanol for 20 min, preincubated with 5% goat serum, and treated with the anti-Ly-6G antibody (1:500, PharMingen) for neutrophils, anti-Mac-3 antibody for macrophages (1: 500, PharMingen), or anti-HO-1 antibody (1:200, SPA 896, StressGen Biotechnologies) for 1 h at 37°C.

    Techniques: Over Expression, Mouse Assay

    HO-1 overexpressing mice are protected from pulmonary hypertension induced by hypoxia. ( a ) RVSP measurements were performed in Tg and non-Tg mice exposed to 2–3 weeks of hypoxia. RVSP did not differ between Tg ( n = 13) and non-Tg ( n = 8) mice at baseline normoxic conditions. Under hypoxia, Tg mice ( n = 25) had significantly lower pressure measurements than non-Tg controls ( n = 8) (*, P

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

    Article Title: Targeted expression of heme oxygenase-1 prevents the pulmonary inflammatory and vascular responses to hypoxia

    doi: 10.1073/pnas.161272598

    Figure Lengend Snippet: HO-1 overexpressing mice are protected from pulmonary hypertension induced by hypoxia. ( a ) RVSP measurements were performed in Tg and non-Tg mice exposed to 2–3 weeks of hypoxia. RVSP did not differ between Tg ( n = 13) and non-Tg ( n = 8) mice at baseline normoxic conditions. Under hypoxia, Tg mice ( n = 25) had significantly lower pressure measurements than non-Tg controls ( n = 8) (*, P

    Article Snippet: Sections were treated with 0.3% hydrogen peroxide in methanol for 20 min, preincubated with 5% goat serum, and treated with the anti-Ly-6G antibody (1:500, PharMingen) for neutrophils, anti-Mac-3 antibody for macrophages (1: 500, PharMingen), or anti-HO-1 antibody (1:200, SPA 896, StressGen Biotechnologies) for 1 h at 37°C.

    Techniques: Mouse Assay

    HO-1 overexpression inhibits hypoxic induction of cytokines in the lung. HO-1 Tg and non-Tg control mice were exposed to hypoxia (8–10% O 2 ) for the times indicated [48 h, 5 days (d), or 2 weeks (w)]. After exposure, total lung RNA (10 μg) was examined for cytokine expression by RNase protection assay. Levels of L32, a ribosomal protein mRNA, were unaffected by hypoxia and served as an internal control. Expression of TNFα 3 h after hypoxia in non-Tg mice is also shown ( Lower ). Similar results were observed in four independent experiments.

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

    Article Title: Targeted expression of heme oxygenase-1 prevents the pulmonary inflammatory and vascular responses to hypoxia

    doi: 10.1073/pnas.161272598

    Figure Lengend Snippet: HO-1 overexpression inhibits hypoxic induction of cytokines in the lung. HO-1 Tg and non-Tg control mice were exposed to hypoxia (8–10% O 2 ) for the times indicated [48 h, 5 days (d), or 2 weeks (w)]. After exposure, total lung RNA (10 μg) was examined for cytokine expression by RNase protection assay. Levels of L32, a ribosomal protein mRNA, were unaffected by hypoxia and served as an internal control. Expression of TNFα 3 h after hypoxia in non-Tg mice is also shown ( Lower ). Similar results were observed in four independent experiments.

    Article Snippet: Sections were treated with 0.3% hydrogen peroxide in methanol for 20 min, preincubated with 5% goat serum, and treated with the anti-Ly-6G antibody (1:500, PharMingen) for neutrophils, anti-Mac-3 antibody for macrophages (1: 500, PharMingen), or anti-HO-1 antibody (1:200, SPA 896, StressGen Biotechnologies) for 1 h at 37°C.

    Techniques: Over Expression, Mouse Assay, Expressing, Rnase Protection Assay

    Expression of HO-1 in transgenic mice. ( a ) Northern blot analysis for HO-1 and HO-2. HO-1 ( Top ) and HO-2 mRNA levels ( Middle ) were examined in total RNA (30 μg) extracted from the lungs of HO-1 transgenic mice (Tg) or nontransgenic littermates (non-Tg). β -actin mRNA levels were examined to confirm equal loading ( Bottom ). This blot is representative of three independent experiments, six mice per group. ( b ) Western blot analysis for HO-1 in the lung. Lung whole-cell lysates (30 μg) from five to six HO-1 Tg and a corresponding number of non-Tg littermates were analyzed for HO-1 protein levels using an anti-HO-1 antibody. The same blot was reprobed with an anti-β-actin antibody to verify equal loading ( Lower ). Note that the anti-HO-1 antibody used here is raised against the N-terminal amino acid sequence of the protein, a region conserved between mouse and human HO-1. ( c ) Western blot analysis for HO-1 in nonlung tissues. Tissue distribution of HO-1 was analyzed in whole cell lysates (30 μg) of heart, liver, spleen, and kidney from Tg mice (+) or non-Tg littermates (−). This blot represents identical patterns of expression observed in three Tg and three non-Tg animals.

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

    Article Title: Targeted expression of heme oxygenase-1 prevents the pulmonary inflammatory and vascular responses to hypoxia

    doi: 10.1073/pnas.161272598

    Figure Lengend Snippet: Expression of HO-1 in transgenic mice. ( a ) Northern blot analysis for HO-1 and HO-2. HO-1 ( Top ) and HO-2 mRNA levels ( Middle ) were examined in total RNA (30 μg) extracted from the lungs of HO-1 transgenic mice (Tg) or nontransgenic littermates (non-Tg). β -actin mRNA levels were examined to confirm equal loading ( Bottom ). This blot is representative of three independent experiments, six mice per group. ( b ) Western blot analysis for HO-1 in the lung. Lung whole-cell lysates (30 μg) from five to six HO-1 Tg and a corresponding number of non-Tg littermates were analyzed for HO-1 protein levels using an anti-HO-1 antibody. The same blot was reprobed with an anti-β-actin antibody to verify equal loading ( Lower ). Note that the anti-HO-1 antibody used here is raised against the N-terminal amino acid sequence of the protein, a region conserved between mouse and human HO-1. ( c ) Western blot analysis for HO-1 in nonlung tissues. Tissue distribution of HO-1 was analyzed in whole cell lysates (30 μg) of heart, liver, spleen, and kidney from Tg mice (+) or non-Tg littermates (−). This blot represents identical patterns of expression observed in three Tg and three non-Tg animals.

    Article Snippet: Sections were treated with 0.3% hydrogen peroxide in methanol for 20 min, preincubated with 5% goat serum, and treated with the anti-Ly-6G antibody (1:500, PharMingen) for neutrophils, anti-Mac-3 antibody for macrophages (1: 500, PharMingen), or anti-HO-1 antibody (1:200, SPA 896, StressGen Biotechnologies) for 1 h at 37°C.

    Techniques: Expressing, Transgenic Assay, Mouse Assay, Northern Blot, Western Blot, Sequencing

    Activation of H-Ras induces HO-1 promoter activity: A and B , Caki-1 and 786-0 cells were co-transfected with the HO-1 promoter-luciferase construct (0.5 μg) and either different amounts (0.5 and 1.0 μg) of active H-Ras overexpression plasmid

    Journal: The Journal of Biological Chemistry

    Article Title: The Heme Oxygenase-1 Protein Is Overexpressed in Human Renal Cancer Cells following Activation of the Ras-Raf-ERK Pathway and Mediates Anti-Apoptotic Signal *

    doi: 10.1074/jbc.M111.248401

    Figure Lengend Snippet: Activation of H-Ras induces HO-1 promoter activity: A and B , Caki-1 and 786-0 cells were co-transfected with the HO-1 promoter-luciferase construct (0.5 μg) and either different amounts (0.5 and 1.0 μg) of active H-Ras overexpression plasmid

    Article Snippet: The membranes were incubated with the following primary antibodies: anti-Ras (BD Transduction Laboratories); anti-HO-1 (R & D systems); anti-H-Ras, anti-Raf-1, anti-Lamin A, and anti-Nrf2 (Santa Cruz Biotechnology); anti-β-actin and anti-GAPDH (Sigma-Aldrich).

    Techniques: Activation Assay, Activity Assay, Transfection, Luciferase, Construct, Over Expression, Plasmid Preparation

    Raf-1 and ERK are important intermediary molecules for H-Ras-induced HO-1 transcription: A and B , 786–0 and Caki-1 cells were pre-treated with RKI/vehicle, and then co-transfected with the HO-1 promoter-luciferase construct (0.5 μg) and

    Journal: The Journal of Biological Chemistry

    Article Title: The Heme Oxygenase-1 Protein Is Overexpressed in Human Renal Cancer Cells following Activation of the Ras-Raf-ERK Pathway and Mediates Anti-Apoptotic Signal *

    doi: 10.1074/jbc.M111.248401

    Figure Lengend Snippet: Raf-1 and ERK are important intermediary molecules for H-Ras-induced HO-1 transcription: A and B , 786–0 and Caki-1 cells were pre-treated with RKI/vehicle, and then co-transfected with the HO-1 promoter-luciferase construct (0.5 μg) and

    Article Snippet: The membranes were incubated with the following primary antibodies: anti-Ras (BD Transduction Laboratories); anti-HO-1 (R & D systems); anti-H-Ras, anti-Raf-1, anti-Lamin A, and anti-Nrf2 (Santa Cruz Biotechnology); anti-β-actin and anti-GAPDH (Sigma-Aldrich).

    Techniques: Transfection, Luciferase, Construct

    Nrf2 is overexpressed in RCC, and plays an important role in HO-1 expression. A , representative photomicrographs show the expression of Nrf2 in human RCC and normal kidney tissues detected by immunohistochemistry. Brown color dots , expression of Nrf2.

    Journal: The Journal of Biological Chemistry

    Article Title: The Heme Oxygenase-1 Protein Is Overexpressed in Human Renal Cancer Cells following Activation of the Ras-Raf-ERK Pathway and Mediates Anti-Apoptotic Signal *

    doi: 10.1074/jbc.M111.248401

    Figure Lengend Snippet: Nrf2 is overexpressed in RCC, and plays an important role in HO-1 expression. A , representative photomicrographs show the expression of Nrf2 in human RCC and normal kidney tissues detected by immunohistochemistry. Brown color dots , expression of Nrf2.

    Article Snippet: The membranes were incubated with the following primary antibodies: anti-Ras (BD Transduction Laboratories); anti-HO-1 (R & D systems); anti-H-Ras, anti-Raf-1, anti-Lamin A, and anti-Nrf2 (Santa Cruz Biotechnology); anti-β-actin and anti-GAPDH (Sigma-Aldrich).

    Techniques: Expressing, Immunohistochemistry

    Activation of H-Ras promotes overexpression of HO-1 mRNA and protein: A and B , 786-0 and Caki-1 cells were transfected with either H-Ras(12V) ( filled column ) or the empty expression vector ( open column ). Following 48 h of transfection, total RNA was isolated

    Journal: The Journal of Biological Chemistry

    Article Title: The Heme Oxygenase-1 Protein Is Overexpressed in Human Renal Cancer Cells following Activation of the Ras-Raf-ERK Pathway and Mediates Anti-Apoptotic Signal *

    doi: 10.1074/jbc.M111.248401

    Figure Lengend Snippet: Activation of H-Ras promotes overexpression of HO-1 mRNA and protein: A and B , 786-0 and Caki-1 cells were transfected with either H-Ras(12V) ( filled column ) or the empty expression vector ( open column ). Following 48 h of transfection, total RNA was isolated

    Article Snippet: The membranes were incubated with the following primary antibodies: anti-Ras (BD Transduction Laboratories); anti-HO-1 (R & D systems); anti-H-Ras, anti-Raf-1, anti-Lamin A, and anti-Nrf2 (Santa Cruz Biotechnology); anti-β-actin and anti-GAPDH (Sigma-Aldrich).

    Techniques: Activation Assay, Over Expression, Transfection, Expressing, Plasmid Preparation, Isolation

    H-Ras-induced survival of renal cancer cells involves HO-1. 786-0 cells were transfected with either HO-1 siRNA (50 n m ) or control siRNA. Following 24 h of siRNA transfection, the cells were transfected with H-Ras(12V) (1.0 μg) or empty vector,

    Journal: The Journal of Biological Chemistry

    Article Title: The Heme Oxygenase-1 Protein Is Overexpressed in Human Renal Cancer Cells following Activation of the Ras-Raf-ERK Pathway and Mediates Anti-Apoptotic Signal *

    doi: 10.1074/jbc.M111.248401

    Figure Lengend Snippet: H-Ras-induced survival of renal cancer cells involves HO-1. 786-0 cells were transfected with either HO-1 siRNA (50 n m ) or control siRNA. Following 24 h of siRNA transfection, the cells were transfected with H-Ras(12V) (1.0 μg) or empty vector,

    Article Snippet: The membranes were incubated with the following primary antibodies: anti-Ras (BD Transduction Laboratories); anti-HO-1 (R & D systems); anti-H-Ras, anti-Raf-1, anti-Lamin A, and anti-Nrf2 (Santa Cruz Biotechnology); anti-β-actin and anti-GAPDH (Sigma-Aldrich).

    Techniques: Transfection, Plasmid Preparation

    Inhibition of ERK induces apoptosis of renal cancer cells, and the overexpression of HO-1 can inhibit this process: 786-0 cells were treated with different combinations of CoPP (10 μ m ) and PD98059 (50 μ m ) for 24 h; control cells were treated

    Journal: The Journal of Biological Chemistry

    Article Title: The Heme Oxygenase-1 Protein Is Overexpressed in Human Renal Cancer Cells following Activation of the Ras-Raf-ERK Pathway and Mediates Anti-Apoptotic Signal *

    doi: 10.1074/jbc.M111.248401

    Figure Lengend Snippet: Inhibition of ERK induces apoptosis of renal cancer cells, and the overexpression of HO-1 can inhibit this process: 786-0 cells were treated with different combinations of CoPP (10 μ m ) and PD98059 (50 μ m ) for 24 h; control cells were treated

    Article Snippet: The membranes were incubated with the following primary antibodies: anti-Ras (BD Transduction Laboratories); anti-HO-1 (R & D systems); anti-H-Ras, anti-Raf-1, anti-Lamin A, and anti-Nrf2 (Santa Cruz Biotechnology); anti-β-actin and anti-GAPDH (Sigma-Aldrich).

    Techniques: Inhibition, Over Expression

    Nrf2 is important for H-Ras- and ERK-induced HO-1 transcription: A , 786-0 cells were transfected (overnight) with either different concentrations of H-Ras(12V) or empty vector ( left panel ); treated with either PD98059 (50 μ m ) or vehicle alone

    Journal: The Journal of Biological Chemistry

    Article Title: The Heme Oxygenase-1 Protein Is Overexpressed in Human Renal Cancer Cells following Activation of the Ras-Raf-ERK Pathway and Mediates Anti-Apoptotic Signal *

    doi: 10.1074/jbc.M111.248401

    Figure Lengend Snippet: Nrf2 is important for H-Ras- and ERK-induced HO-1 transcription: A , 786-0 cells were transfected (overnight) with either different concentrations of H-Ras(12V) or empty vector ( left panel ); treated with either PD98059 (50 μ m ) or vehicle alone

    Article Snippet: The membranes were incubated with the following primary antibodies: anti-Ras (BD Transduction Laboratories); anti-HO-1 (R & D systems); anti-H-Ras, anti-Raf-1, anti-Lamin A, and anti-Nrf2 (Santa Cruz Biotechnology); anti-β-actin and anti-GAPDH (Sigma-Aldrich).

    Techniques: Transfection, Plasmid Preparation

    H-Ras promotes HO-1 transcriptional activation primarily through the Raf signaling pathway: A and B , 786–0 and Caki-1 cells were transfected with the HO-1 promoter-luciferase construct (0.5 μg) and either 0.5 μg of an effector

    Journal: The Journal of Biological Chemistry

    Article Title: The Heme Oxygenase-1 Protein Is Overexpressed in Human Renal Cancer Cells following Activation of the Ras-Raf-ERK Pathway and Mediates Anti-Apoptotic Signal *

    doi: 10.1074/jbc.M111.248401

    Figure Lengend Snippet: H-Ras promotes HO-1 transcriptional activation primarily through the Raf signaling pathway: A and B , 786–0 and Caki-1 cells were transfected with the HO-1 promoter-luciferase construct (0.5 μg) and either 0.5 μg of an effector

    Article Snippet: The membranes were incubated with the following primary antibodies: anti-Ras (BD Transduction Laboratories); anti-HO-1 (R & D systems); anti-H-Ras, anti-Raf-1, anti-Lamin A, and anti-Nrf2 (Santa Cruz Biotechnology); anti-β-actin and anti-GAPDH (Sigma-Aldrich).

    Techniques: Activation Assay, Transfection, Luciferase, Construct

    Effect of lucidone on HO-1 expression. (A to C) Concentration-dependent induction of HO-1 promoter activity (A), RNA transcription (B), and protein synthesis by lucidone (C). For the promoter activity assay, Ava5 cells were transiently transfected with

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Lucidone Suppresses Hepatitis C Virus Replication by Nrf2-Mediated Heme Oxygenase-1 Induction

    doi: 10.1128/AAC.02053-12

    Figure Lengend Snippet: Effect of lucidone on HO-1 expression. (A to C) Concentration-dependent induction of HO-1 promoter activity (A), RNA transcription (B), and protein synthesis by lucidone (C). For the promoter activity assay, Ava5 cells were transiently transfected with

    Article Snippet: Membranes were probed with either anti-NS5B (1:5,000; Abcam, Cambridge, MA), anti-GAPDH (1:10,000; GeneTex, CA), anti-HO-1 (1:3,000; GeneTex), anti-BVR (1:1,000; Abcam, Cambridge, MA), or anti-Nrf2 (1:3,000; GeneTex) antibody.

    Techniques: Expressing, Concentration Assay, Activity Assay, Transfection

    Induction of the antiviral IFN responses by lucidone in HCV replicon cells. (A) Concentration-dependent induction of ISRE activity by lucidone. (B) Restoration of lucidone-induced HO-1 activity by HO-1 inhibitor SnPP. (C to E) Concentration-dependent

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Lucidone Suppresses Hepatitis C Virus Replication by Nrf2-Mediated Heme Oxygenase-1 Induction

    doi: 10.1128/AAC.02053-12

    Figure Lengend Snippet: Induction of the antiviral IFN responses by lucidone in HCV replicon cells. (A) Concentration-dependent induction of ISRE activity by lucidone. (B) Restoration of lucidone-induced HO-1 activity by HO-1 inhibitor SnPP. (C to E) Concentration-dependent

    Article Snippet: Membranes were probed with either anti-NS5B (1:5,000; Abcam, Cambridge, MA), anti-GAPDH (1:10,000; GeneTex, CA), anti-HO-1 (1:3,000; GeneTex), anti-BVR (1:1,000; Abcam, Cambridge, MA), or anti-Nrf2 (1:3,000; GeneTex) antibody.

    Techniques: Concentration Assay, Activity Assay

    Restoration of lucidone-suppressed HCV protein synthesis and RNA replication by HO-1 inhibition. Ava5 cells were treated with or without different concentrations (0 to 20 μM) of the HO-1 specific inhibitor SnPP in the presence of 30 μM

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Lucidone Suppresses Hepatitis C Virus Replication by Nrf2-Mediated Heme Oxygenase-1 Induction

    doi: 10.1128/AAC.02053-12

    Figure Lengend Snippet: Restoration of lucidone-suppressed HCV protein synthesis and RNA replication by HO-1 inhibition. Ava5 cells were treated with or without different concentrations (0 to 20 μM) of the HO-1 specific inhibitor SnPP in the presence of 30 μM

    Article Snippet: Membranes were probed with either anti-NS5B (1:5,000; Abcam, Cambridge, MA), anti-GAPDH (1:10,000; GeneTex, CA), anti-HO-1 (1:3,000; GeneTex), anti-BVR (1:1,000; Abcam, Cambridge, MA), or anti-Nrf2 (1:3,000; GeneTex) antibody.

    Techniques: Inhibition

    Effect of SIN on oxidative stress. After treatment with 0.25, 0.5, or 1 mM for 24 h, the level of MDA ( A ) and activity of SOD ( B ) and CAT ( C ) was detected; Protein ( D ) and mRNA ( E ) levels of Nrf2, HO-1, and NQO-1 were detected using Western blotting and qRT-PCR, respectively. Controls: cells without any treatment; BK: 1 μg/ml BK treatment group; BK + SIN 0.25: 1 μg/ml BK + 0.25 nM SIN treatment group; BK + SIN 0.5: 1 μg/ml BK + 0.5 nM SIN treatment group; BK + SIN 1: 1 μg/ml BK + 1 nM SIN treatment group; * p

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

    Article Title: Sinomenine Regulates Inflammatory Response and Oxidative Stress via Nuclear Factor kappa B (NF-κB) and NF-E2-Related Factor 2 (Nrf2) Signaling Pathways in Ankle Fractures in Children

    doi: 10.12659/MSM.910740

    Figure Lengend Snippet: Effect of SIN on oxidative stress. After treatment with 0.25, 0.5, or 1 mM for 24 h, the level of MDA ( A ) and activity of SOD ( B ) and CAT ( C ) was detected; Protein ( D ) and mRNA ( E ) levels of Nrf2, HO-1, and NQO-1 were detected using Western blotting and qRT-PCR, respectively. Controls: cells without any treatment; BK: 1 μg/ml BK treatment group; BK + SIN 0.25: 1 μg/ml BK + 0.25 nM SIN treatment group; BK + SIN 0.5: 1 μg/ml BK + 0.5 nM SIN treatment group; BK + SIN 1: 1 μg/ml BK + 1 nM SIN treatment group; * p

    Article Snippet: We obtained anti-Nrf2 from Santa Cruz Biotechnology (USA), anti-HO-1 was purchased from Enzo Life Sciences (USA), and anti-NQO-1 was obtained from Novus biological (USA).

    Techniques: Multiple Displacement Amplification, Activity Assay, Western Blot, Quantitative RT-PCR

    CORM-3-induced HO-1 expression is mediated through p42/p44 mitogen-activated protein kinase (MAPK). (A) RBA-1 cells were pretreated with various concentrations of U0126 for 1 h, and then incubated with 30 μM CORM-3 for 6 h. The levels of HO-1 and GAPDH (as an internal control) protein expression were determined by Western blot. (B) RBA-1 cells were pretreated with 10 μM U0126 for 1 h and then incubated with 30 μM CORM-3 for 4 h. The levels of HO-1 mRNA were determined by real-time PCR (Open bars). Cells were transiently transfected with HO-1 report gene together a β-galactosidase plasmid, pretreated with U0126 (10 μM) for 1 h, and then incubated with CORM-3 for 1 h. Promoter activity was determined in the cell lysates (Gray bars). (C) RBA-1 cells were transfected with p44 or p42 siRNA and then incubated with 30 μM CORM-3 for 6 h. The levels of total p42, p44 and HO-1 were determined by Western blot. (D) RBA-1 cells were pretreated with 10 μM U0126, 10 μM SU6656, 10 μM PF431396, or 3 μM Gö6983, for 1 h and then incubated with 30 μM CORM-3 for the indicated time intervals. Phosphorylation of p42/p44 MAPK was determined by Western blot using an anti-phospho-p42/p44 MAPK or anti-p42/p44 MAPK antibody. Data are expressed as the mean ± SEM of three independent experiments ( n = 3). # p

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Heme Oxygenase-1 Induction by Carbon Monoxide Releasing Molecule-3 Suppresses Interleukin-1β-Mediated Neuroinflammation

    doi: 10.3389/fnmol.2017.00387

    Figure Lengend Snippet: CORM-3-induced HO-1 expression is mediated through p42/p44 mitogen-activated protein kinase (MAPK). (A) RBA-1 cells were pretreated with various concentrations of U0126 for 1 h, and then incubated with 30 μM CORM-3 for 6 h. The levels of HO-1 and GAPDH (as an internal control) protein expression were determined by Western blot. (B) RBA-1 cells were pretreated with 10 μM U0126 for 1 h and then incubated with 30 μM CORM-3 for 4 h. The levels of HO-1 mRNA were determined by real-time PCR (Open bars). Cells were transiently transfected with HO-1 report gene together a β-galactosidase plasmid, pretreated with U0126 (10 μM) for 1 h, and then incubated with CORM-3 for 1 h. Promoter activity was determined in the cell lysates (Gray bars). (C) RBA-1 cells were transfected with p44 or p42 siRNA and then incubated with 30 μM CORM-3 for 6 h. The levels of total p42, p44 and HO-1 were determined by Western blot. (D) RBA-1 cells were pretreated with 10 μM U0126, 10 μM SU6656, 10 μM PF431396, or 3 μM Gö6983, for 1 h and then incubated with 30 μM CORM-3 for the indicated time intervals. Phosphorylation of p42/p44 MAPK was determined by Western blot using an anti-phospho-p42/p44 MAPK or anti-p42/p44 MAPK antibody. Data are expressed as the mean ± SEM of three independent experiments ( n = 3). # p

    Article Snippet: Anti-HO-1 antibody (ADI-SPA-895), actinomycin D (ActD), cycloheximide (CHI), PP1, SU6656, PF431396, Gö6976, Gö6983, Ro31-8220, U0126 and Tanshinone IIA (TSIIA) were from Enzo Life Sciences (Farmingdale, NY, USA).

    Techniques: Expressing, Incubation, Western Blot, Real-time Polymerase Chain Reaction, Transfection, Plasmid Preparation, Activity Assay

    Schematic representation of signaling pathways involved in CORM-3-induced HO-1 expression and protected against IL-1β-induced cell migration in RBA-1 cells. CORM-3 activated c-Src/Pyk2/PKCα/p42/p44 MAPK/AP-1 pathway to induce HO-1 expression which suppressed the IL-1β-induced MMP-9 mRNA expression and cell migration.

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Heme Oxygenase-1 Induction by Carbon Monoxide Releasing Molecule-3 Suppresses Interleukin-1β-Mediated Neuroinflammation

    doi: 10.3389/fnmol.2017.00387

    Figure Lengend Snippet: Schematic representation of signaling pathways involved in CORM-3-induced HO-1 expression and protected against IL-1β-induced cell migration in RBA-1 cells. CORM-3 activated c-Src/Pyk2/PKCα/p42/p44 MAPK/AP-1 pathway to induce HO-1 expression which suppressed the IL-1β-induced MMP-9 mRNA expression and cell migration.

    Article Snippet: Anti-HO-1 antibody (ADI-SPA-895), actinomycin D (ActD), cycloheximide (CHI), PP1, SU6656, PF431396, Gö6976, Gö6983, Ro31-8220, U0126 and Tanshinone IIA (TSIIA) were from Enzo Life Sciences (Farmingdale, NY, USA).

    Techniques: Expressing, Migration

    CORM-3-induced HO-1 expression reduces the interleukin (IL)-1β-induced matrix metalloproteinase-9 (MMP-9) expression and cell migration. (A,B) RBA-1 cells were incubated with pretreated with (30 μM) CORM-3 for 4 h, and then incubated with IL-1β (10 μg/ml) for 6 h. The levels of HO-1 and MMP-9 mRNA were determined by real-time PCR. (C) RBA-1 cells were pretreated with (10 μM) MMP2/9i for 1 h or transfected HO-1 siRNA followed by (30 μM) CORM-3, and infected with adenovirus-introduced HO-1, and then incubated with IL-1β (10 μg/ml) for 48 h in the presence of (1 μM) hydroxyurea. Phase contrast images of cell migration were taken at 48 h. Data are expressed as the mean ± SEM of three independent experiments ( n = 3). # p

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Heme Oxygenase-1 Induction by Carbon Monoxide Releasing Molecule-3 Suppresses Interleukin-1β-Mediated Neuroinflammation

    doi: 10.3389/fnmol.2017.00387

    Figure Lengend Snippet: CORM-3-induced HO-1 expression reduces the interleukin (IL)-1β-induced matrix metalloproteinase-9 (MMP-9) expression and cell migration. (A,B) RBA-1 cells were incubated with pretreated with (30 μM) CORM-3 for 4 h, and then incubated with IL-1β (10 μg/ml) for 6 h. The levels of HO-1 and MMP-9 mRNA were determined by real-time PCR. (C) RBA-1 cells were pretreated with (10 μM) MMP2/9i for 1 h or transfected HO-1 siRNA followed by (30 μM) CORM-3, and infected with adenovirus-introduced HO-1, and then incubated with IL-1β (10 μg/ml) for 48 h in the presence of (1 μM) hydroxyurea. Phase contrast images of cell migration were taken at 48 h. Data are expressed as the mean ± SEM of three independent experiments ( n = 3). # p

    Article Snippet: Anti-HO-1 antibody (ADI-SPA-895), actinomycin D (ActD), cycloheximide (CHI), PP1, SU6656, PF431396, Gö6976, Gö6983, Ro31-8220, U0126 and Tanshinone IIA (TSIIA) were from Enzo Life Sciences (Farmingdale, NY, USA).

    Techniques: Expressing, Migration, Incubation, Real-time Polymerase Chain Reaction, Transfection, Infection

    CORM-3 induces HO-1 expression via transcription and translation. RBA-1 cells were pretreated with various concentrations of either (A) actinomycin D (ActD) or (B) cycloheximide (CHI) for 1 h and then incubated with 30 μM CORM-3 for 6 h. The levels of HO-1 and GAPDH (as an internal control) protein expression were determined by Western blot. (C) RBA-1 cells were pretreated with 100 nM ActD for 1 h and then incubated with 30 μM CORM-3 for 4 h. The levels of HO-1 mRNA were determined by real-time PCR (open bars). Cells were transiently transfected with HO-1 report gene together a β-galactosidase plasmid, pretreated with 100 nM ActD for 1 h, and then incubated with 30 μM CORM-3 for 1 h. Promoter activity was determined in the cell lysates (shaded bars). Data are expressed as the mean ± SEM of three independent experiments ( n = 3). # p

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Heme Oxygenase-1 Induction by Carbon Monoxide Releasing Molecule-3 Suppresses Interleukin-1β-Mediated Neuroinflammation

    doi: 10.3389/fnmol.2017.00387

    Figure Lengend Snippet: CORM-3 induces HO-1 expression via transcription and translation. RBA-1 cells were pretreated with various concentrations of either (A) actinomycin D (ActD) or (B) cycloheximide (CHI) for 1 h and then incubated with 30 μM CORM-3 for 6 h. The levels of HO-1 and GAPDH (as an internal control) protein expression were determined by Western blot. (C) RBA-1 cells were pretreated with 100 nM ActD for 1 h and then incubated with 30 μM CORM-3 for 4 h. The levels of HO-1 mRNA were determined by real-time PCR (open bars). Cells were transiently transfected with HO-1 report gene together a β-galactosidase plasmid, pretreated with 100 nM ActD for 1 h, and then incubated with 30 μM CORM-3 for 1 h. Promoter activity was determined in the cell lysates (shaded bars). Data are expressed as the mean ± SEM of three independent experiments ( n = 3). # p

    Article Snippet: Anti-HO-1 antibody (ADI-SPA-895), actinomycin D (ActD), cycloheximide (CHI), PP1, SU6656, PF431396, Gö6976, Gö6983, Ro31-8220, U0126 and Tanshinone IIA (TSIIA) were from Enzo Life Sciences (Farmingdale, NY, USA).

    Techniques: Expressing, Incubation, Western Blot, Real-time Polymerase Chain Reaction, Transfection, Plasmid Preparation, Activity Assay

    CORM-3 induces HO-1 expression via c-Src and Pyk2. (A) RBA-1 cells were pretreated with various concentrations of PP1, SU6656 or PF431396 for 1 h, and then incubated with 30 μM CORM-3 for 6 h. The levels of HO-1 and GAPDH (as an internal control) protein expression were determined by Western blot. (B) RBA-1 cells were pretreated with 10 μM PP1, 10 μM SU6656, or 10 μM PF431396 for 1 h and then incubated with 30 μM CORM-3 for 4 h. The levels of HO-1 mRNA were determined by real-time PCR (open bars). Cells were transiently transfected with HO-1 report gene together a β-galactosidase plasmid, pretreated with 10 μM PP1, 10 μM SU6656, or 10 μM PF431396 for 1 h, and then incubated with 30 μM CORM-3 for 1 h. Promoter activity was determined in the cell lysates (shaded bars). (C) RBA-1 cells were pretreated without or with 10 μM PP1, 10 μM SU6656, or 10 μM PF431396 for 1 h and then incubated with 30 μM CORM-3 for the indicated time intervals. The cell lysates were subjected to western blot using an anti-phospho-c-Src, anti-phospho-Pyk2, anti-c-Src, or anti-Pyk2 antibody. (D) Cells were transfected with c-Src siRNA or Pyk2 siRNA and then incubated with 30 μM CORM-3 for 6 h. The cell lysates were subjected to western blot using an anti-HO-1, anti-c-Src, anti-Pyk2 or GAPDH (as internal control) antibody. Data were expressed as mean ± SEM of three independent experiments ( n = 3). * p

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Heme Oxygenase-1 Induction by Carbon Monoxide Releasing Molecule-3 Suppresses Interleukin-1β-Mediated Neuroinflammation

    doi: 10.3389/fnmol.2017.00387

    Figure Lengend Snippet: CORM-3 induces HO-1 expression via c-Src and Pyk2. (A) RBA-1 cells were pretreated with various concentrations of PP1, SU6656 or PF431396 for 1 h, and then incubated with 30 μM CORM-3 for 6 h. The levels of HO-1 and GAPDH (as an internal control) protein expression were determined by Western blot. (B) RBA-1 cells were pretreated with 10 μM PP1, 10 μM SU6656, or 10 μM PF431396 for 1 h and then incubated with 30 μM CORM-3 for 4 h. The levels of HO-1 mRNA were determined by real-time PCR (open bars). Cells were transiently transfected with HO-1 report gene together a β-galactosidase plasmid, pretreated with 10 μM PP1, 10 μM SU6656, or 10 μM PF431396 for 1 h, and then incubated with 30 μM CORM-3 for 1 h. Promoter activity was determined in the cell lysates (shaded bars). (C) RBA-1 cells were pretreated without or with 10 μM PP1, 10 μM SU6656, or 10 μM PF431396 for 1 h and then incubated with 30 μM CORM-3 for the indicated time intervals. The cell lysates were subjected to western blot using an anti-phospho-c-Src, anti-phospho-Pyk2, anti-c-Src, or anti-Pyk2 antibody. (D) Cells were transfected with c-Src siRNA or Pyk2 siRNA and then incubated with 30 μM CORM-3 for 6 h. The cell lysates were subjected to western blot using an anti-HO-1, anti-c-Src, anti-Pyk2 or GAPDH (as internal control) antibody. Data were expressed as mean ± SEM of three independent experiments ( n = 3). * p

    Article Snippet: Anti-HO-1 antibody (ADI-SPA-895), actinomycin D (ActD), cycloheximide (CHI), PP1, SU6656, PF431396, Gö6976, Gö6983, Ro31-8220, U0126 and Tanshinone IIA (TSIIA) were from Enzo Life Sciences (Farmingdale, NY, USA).

    Techniques: Expressing, Incubation, Western Blot, Real-time Polymerase Chain Reaction, Transfection, Plasmid Preparation, Activity Assay

    CORM-3-induced HO-1 protein expression is mediated via protein kinase C (PKCα). (A) RBA-1 cells were pretreated with various concentrations of Ro31-8220, Gö6976, or Gö6983 for 1 h, and then incubated with 30 μM CORM-3 for 6 h. The levels of HO-1 and GAPDH (as an internal control) protein expression were determined by Western blot. (B) RBA-1 cells were pretreated with 3 μM Gö6976, 3 μM Gö6983, or 3 μM Ro31-8220 for 1 h and then incubated with 30 μM CORM-3 for 4 h. The levels of HO-1 mRNA were determined by real-time PCR (Open bars). Cells were transiently transfected with HO-1 report gene together a β-galactosidase plasmid, pretreated with 3 μM Gö6976, 3 μM Gö6983, or 3 μM Ro31-8220 for 1 h, and then incubated with 30 μM CORM-3 for 1 h. Promoter activity was determined in the cell lysates (Gray bars). (C) RBA-1 cells were transfected with PKCα siRNA and then incubated with 30 μM CORM-3 for 6 h. The cell lysates were subjected to western blot using an anti-HO-1, anti-PKCα, or anti-GAPDH (as internal control). The densitometric quantifications are indicated on the panels. (D) RBA-1 cells were pretreated with 3 μM Ro31-8220, 3 μM Gö6983, 3 μM Gö6976, 10 μM SU6656, or 10 μM PF431396 for 1 h, and stimulated with 30 μM CORM-3 for the indicated time intervals. Phosphorylation of PKCα/βII was determined by Western blot using an anti-phospho-PKCα/βII or anti-PKCα antibody. Data are expressed as the mean ± SEM of three independent experiments ( n = 3). * p

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Heme Oxygenase-1 Induction by Carbon Monoxide Releasing Molecule-3 Suppresses Interleukin-1β-Mediated Neuroinflammation

    doi: 10.3389/fnmol.2017.00387

    Figure Lengend Snippet: CORM-3-induced HO-1 protein expression is mediated via protein kinase C (PKCα). (A) RBA-1 cells were pretreated with various concentrations of Ro31-8220, Gö6976, or Gö6983 for 1 h, and then incubated with 30 μM CORM-3 for 6 h. The levels of HO-1 and GAPDH (as an internal control) protein expression were determined by Western blot. (B) RBA-1 cells were pretreated with 3 μM Gö6976, 3 μM Gö6983, or 3 μM Ro31-8220 for 1 h and then incubated with 30 μM CORM-3 for 4 h. The levels of HO-1 mRNA were determined by real-time PCR (Open bars). Cells were transiently transfected with HO-1 report gene together a β-galactosidase plasmid, pretreated with 3 μM Gö6976, 3 μM Gö6983, or 3 μM Ro31-8220 for 1 h, and then incubated with 30 μM CORM-3 for 1 h. Promoter activity was determined in the cell lysates (Gray bars). (C) RBA-1 cells were transfected with PKCα siRNA and then incubated with 30 μM CORM-3 for 6 h. The cell lysates were subjected to western blot using an anti-HO-1, anti-PKCα, or anti-GAPDH (as internal control). The densitometric quantifications are indicated on the panels. (D) RBA-1 cells were pretreated with 3 μM Ro31-8220, 3 μM Gö6983, 3 μM Gö6976, 10 μM SU6656, or 10 μM PF431396 for 1 h, and stimulated with 30 μM CORM-3 for the indicated time intervals. Phosphorylation of PKCα/βII was determined by Western blot using an anti-phospho-PKCα/βII or anti-PKCα antibody. Data are expressed as the mean ± SEM of three independent experiments ( n = 3). * p

    Article Snippet: Anti-HO-1 antibody (ADI-SPA-895), actinomycin D (ActD), cycloheximide (CHI), PP1, SU6656, PF431396, Gö6976, Gö6983, Ro31-8220, U0126 and Tanshinone IIA (TSIIA) were from Enzo Life Sciences (Farmingdale, NY, USA).

    Techniques: Expressing, Incubation, Western Blot, Real-time Polymerase Chain Reaction, Transfection, Plasmid Preparation, Activity Assay

    CORM-3-induced HO-1 expression is mediated through activator protein (AP)-1. (A) RBA-1 cells were pretreated with various concentrations of Tanshinone IIA (TSIIA) for 1 h, and then stimulated with 30 μM CORM-3 for 6 h. The levels of HO-1 and GAPDH (as an internal control) protein expression were determined by Western blot. (B) RBA-1 cells were pretreated with TSIIA (1 μM) for 1 h and then incubated with 30 μM CORM-3 for 4 h. The levels of HO-1 mRNA were determined by real-time PCR (Open bars). Cells were transiently transfected with HO-1 report gene together a β-galactosidase plasmid, pretreated with TSIIA (1 μM) for 1 h, and then incubated with CORM-3 for 1 h. Promoter activity was determined in the cell lysates (Gray bars). (C) RBA-1 cells were transfected with c-Fos or c-Jun siRNA and then incubated with 30 μM CORM-3 for 6 h. The levels of total protein c-Fos, c-Jun and HO-1 were determined by Western blot. (D) RBA-1 cells were incubated with 30 μM CORM-3 for the indicated time intervals. (E,F) Cells were pretreated without or with (E) PP1 (10 μM), SU6656 (10 μM), or PF431396 (10 μM) and (F) (3 μM) Ro-318220, (3 μM) Gö6976, (3 μM) Gö6983, or (10 μM) U0126 for 1 h and then incubated with (30 μM) CORM-3 for 15 min. (D–F) The levels of c-jun (Open bars) and c-fos (Gray bars) mRNA were determined by real-time PCR. Data are expressed as the mean ± SEM of three independent experiments ( n = 3). * p

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Heme Oxygenase-1 Induction by Carbon Monoxide Releasing Molecule-3 Suppresses Interleukin-1β-Mediated Neuroinflammation

    doi: 10.3389/fnmol.2017.00387

    Figure Lengend Snippet: CORM-3-induced HO-1 expression is mediated through activator protein (AP)-1. (A) RBA-1 cells were pretreated with various concentrations of Tanshinone IIA (TSIIA) for 1 h, and then stimulated with 30 μM CORM-3 for 6 h. The levels of HO-1 and GAPDH (as an internal control) protein expression were determined by Western blot. (B) RBA-1 cells were pretreated with TSIIA (1 μM) for 1 h and then incubated with 30 μM CORM-3 for 4 h. The levels of HO-1 mRNA were determined by real-time PCR (Open bars). Cells were transiently transfected with HO-1 report gene together a β-galactosidase plasmid, pretreated with TSIIA (1 μM) for 1 h, and then incubated with CORM-3 for 1 h. Promoter activity was determined in the cell lysates (Gray bars). (C) RBA-1 cells were transfected with c-Fos or c-Jun siRNA and then incubated with 30 μM CORM-3 for 6 h. The levels of total protein c-Fos, c-Jun and HO-1 were determined by Western blot. (D) RBA-1 cells were incubated with 30 μM CORM-3 for the indicated time intervals. (E,F) Cells were pretreated without or with (E) PP1 (10 μM), SU6656 (10 μM), or PF431396 (10 μM) and (F) (3 μM) Ro-318220, (3 μM) Gö6976, (3 μM) Gö6983, or (10 μM) U0126 for 1 h and then incubated with (30 μM) CORM-3 for 15 min. (D–F) The levels of c-jun (Open bars) and c-fos (Gray bars) mRNA were determined by real-time PCR. Data are expressed as the mean ± SEM of three independent experiments ( n = 3). * p

    Article Snippet: Anti-HO-1 antibody (ADI-SPA-895), actinomycin D (ActD), cycloheximide (CHI), PP1, SU6656, PF431396, Gö6976, Gö6983, Ro31-8220, U0126 and Tanshinone IIA (TSIIA) were from Enzo Life Sciences (Farmingdale, NY, USA).

    Techniques: Expressing, Western Blot, Incubation, Real-time Polymerase Chain Reaction, Transfection, Plasmid Preparation, Activity Assay

    CO-releasing molecule-3 (CORM-3) induces heme oxygenase-1 (HO-1) expression. (A) Rat brain astrocytes (RBA)-1 cells were treated with various concentrations of CORM-3 for the indicated time intervals. The levels of HO-1 and GAPDH (as an internal control) protein expression were determined by Western blot. (B) Cells were treated with 30 μM CORM-3 for the indicated time intervals. The HO-1 mRNA levels were determined by real-time PCR. (C) Cells were co-transfected with HO-1 promoter and β-galactosidase plasmids, and then incubated with 30 μM CORM-3 for the indicated time intervals. HO-1 promoter luciferase activity was determined in the cell lysates. Data were expressed as mean ± standard errors of the mean (SEM) of three independent experiments ( n = 3). * p

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Heme Oxygenase-1 Induction by Carbon Monoxide Releasing Molecule-3 Suppresses Interleukin-1β-Mediated Neuroinflammation

    doi: 10.3389/fnmol.2017.00387

    Figure Lengend Snippet: CO-releasing molecule-3 (CORM-3) induces heme oxygenase-1 (HO-1) expression. (A) Rat brain astrocytes (RBA)-1 cells were treated with various concentrations of CORM-3 for the indicated time intervals. The levels of HO-1 and GAPDH (as an internal control) protein expression were determined by Western blot. (B) Cells were treated with 30 μM CORM-3 for the indicated time intervals. The HO-1 mRNA levels were determined by real-time PCR. (C) Cells were co-transfected with HO-1 promoter and β-galactosidase plasmids, and then incubated with 30 μM CORM-3 for the indicated time intervals. HO-1 promoter luciferase activity was determined in the cell lysates. Data were expressed as mean ± standard errors of the mean (SEM) of three independent experiments ( n = 3). * p

    Article Snippet: Anti-HO-1 antibody (ADI-SPA-895), actinomycin D (ActD), cycloheximide (CHI), PP1, SU6656, PF431396, Gö6976, Gö6983, Ro31-8220, U0126 and Tanshinone IIA (TSIIA) were from Enzo Life Sciences (Farmingdale, NY, USA).

    Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Transfection, Incubation, Luciferase, Activity Assay

    Supernatant of platelets treated with plasma-derived factor VIII products promote the formation of macrophages that are unable to upregulate HO-1 in response to hemoglobin. Human monocytes were cultured to macrophages in the presence of M-CSF for 3 days then for 3 days with M-CSF ±CXCL4 or supernatant from untreated platelets or platelets treated with different factor VIII products or with ADP plus U46619, as indicated. Cells were then incubated with autologous hemoglobin for 18 h, harvested, permeabilized and stained with anti-HO-1-PE prior to analysis by flow cytometry. Data show mean ± standard deviation (SD) from 3 independent experiments. ***p

    Journal: Transfusion Medicine and Hemotherapy

    Article Title: Components in Plasma-Derived Factor VIII, But Not in Recombinant Factor VIII Downregulate Anti-Inflammatory Surface Marker CD163 in Human Macrophages through Release of CXCL4 (Platelet Factor 4)

    doi: 10.1159/000472157

    Figure Lengend Snippet: Supernatant of platelets treated with plasma-derived factor VIII products promote the formation of macrophages that are unable to upregulate HO-1 in response to hemoglobin. Human monocytes were cultured to macrophages in the presence of M-CSF for 3 days then for 3 days with M-CSF ±CXCL4 or supernatant from untreated platelets or platelets treated with different factor VIII products or with ADP plus U46619, as indicated. Cells were then incubated with autologous hemoglobin for 18 h, harvested, permeabilized and stained with anti-HO-1-PE prior to analysis by flow cytometry. Data show mean ± standard deviation (SD) from 3 independent experiments. ***p

    Article Snippet: For intracellular staining of HO-1, cells were fixed with 2% formaldehyde in PBS, permeabilized with 0.1% saponin (Sigma Aldrich) and 0.5% bovine serum albumin (Calbiochem, San Diego, CA, USA) in PBS and subsequently stained with predefined saturating concentrations of anti-HO-1-PE (clone HO-1-2; Enzo Life Sciences, Lörrach, Germany).

    Techniques: Derivative Assay, Cell Culture, Incubation, Staining, Flow Cytometry, Cytometry, Standard Deviation

    Flaccidoside II downregulates the expression and activation of antiapoptotic HO-1, which is involved in the apoptotic facilitation of flaccidoside II. Notes: ( A ) The ST88-14 and S462 cells were treated with either 40.0 μmol L −1 flaccidoside II or DMSO for 48 hours, and the protein expression level of HO-1 was measured by Western blotting, which was normalized to GAPDH. ( B ) The HO activity was measured in ST88-14 and S462 cells at 48 hours after treatment of these cells with 40.0 μmol L −1 flaccidoside II or DMSO. ( C ) The ST88-14 and S462 cells were treated with 40.0 μmol L −1 flaccidoside II with the transfection of either vector plasmid as a negative control or full-length HO-1 plasmid. The expression of HO-1, C-caspase 3, total PARP, and cleaved PARP were determined by Western blotting, and GAPDH was used as loading control ( C ). ( D ) Quantitative analysis of the apoptosis markers cleaved caspase 3 and cleaved PARP in ( C ). ( E ) The apoptotic rates of ST88-14 and S462 cells were determined by Annexin V-PI staining and flow cytometry for quantification. Each bar represents the mean ± SD of four independent experiments. *** P

    Journal: OncoTargets and therapy

    Article Title: Triterpenoid saponin flaccidoside II from Anemone flaccida triggers apoptosis of NF1-associated malignant peripheral nerve sheath tumors via the MAPK-HO-1 pathway

    doi: 10.2147/OTT.S95597

    Figure Lengend Snippet: Flaccidoside II downregulates the expression and activation of antiapoptotic HO-1, which is involved in the apoptotic facilitation of flaccidoside II. Notes: ( A ) The ST88-14 and S462 cells were treated with either 40.0 μmol L −1 flaccidoside II or DMSO for 48 hours, and the protein expression level of HO-1 was measured by Western blotting, which was normalized to GAPDH. ( B ) The HO activity was measured in ST88-14 and S462 cells at 48 hours after treatment of these cells with 40.0 μmol L −1 flaccidoside II or DMSO. ( C ) The ST88-14 and S462 cells were treated with 40.0 μmol L −1 flaccidoside II with the transfection of either vector plasmid as a negative control or full-length HO-1 plasmid. The expression of HO-1, C-caspase 3, total PARP, and cleaved PARP were determined by Western blotting, and GAPDH was used as loading control ( C ). ( D ) Quantitative analysis of the apoptosis markers cleaved caspase 3 and cleaved PARP in ( C ). ( E ) The apoptotic rates of ST88-14 and S462 cells were determined by Annexin V-PI staining and flow cytometry for quantification. Each bar represents the mean ± SD of four independent experiments. *** P

    Article Snippet: The following antibodies were used: anti-HO-1 (Epitomics, Burlingame, CA, USA), antiphospho-p38 (Cell Signaling Technology, Beverly, MA, USA), antiphospho-ERK-1/2 (Cell Signaling Technology), antiphospho-JNK (Santa Cruz Biotechnology Inc., Dallas, TX, USA), anti-p38 (Cell Signaling Technology), anti-ERK-1/2 (Cell Signaling Technology), anti-JNK (Santa Cruz Biotechnology Inc.), and anti-GAPDH (Abcam, Cambridge, MA, USA).

    Techniques: Expressing, Activation Assay, Western Blot, Activity Assay, Transfection, Plasmid Preparation, Negative Control, Staining, Flow Cytometry, Cytometry

    HO-1 downregulation induced by flaccidoside II is mediated by ERK-1/2 and p38 MAPK pathways. Notes: ( A – C ) The ST88-14 and S462 cells were treated with 40.0 μmol L −1 flaccidoside II with performance of ERK-1/2 inhibitors PD98059 or p38 inhibitor SB203580, and the protein expression level of HO-1, C-caspase 3, total PARP, and cleaved PARP were measured by Western blotting, and β-actin was used as loading control ( A ). ( B ) Quantitative analysis of the blots in ( A ). ( C ) The apoptotic rates of cells were determined by Annexin V-PI staining and flow cytometry for quantification. ( D ) HO activity was measured in ST88-14 and S462 cells at 48 hours treatment of 40.0 μmol L −1 flaccidoside II with/without the performance of ERK-1/2 inhibitors PD98059 or p38 inhibitor SB203580, respectively. Each bar represents the mean ± SD of at least four independent experiments. * P

    Journal: OncoTargets and therapy

    Article Title: Triterpenoid saponin flaccidoside II from Anemone flaccida triggers apoptosis of NF1-associated malignant peripheral nerve sheath tumors via the MAPK-HO-1 pathway

    doi: 10.2147/OTT.S95597

    Figure Lengend Snippet: HO-1 downregulation induced by flaccidoside II is mediated by ERK-1/2 and p38 MAPK pathways. Notes: ( A – C ) The ST88-14 and S462 cells were treated with 40.0 μmol L −1 flaccidoside II with performance of ERK-1/2 inhibitors PD98059 or p38 inhibitor SB203580, and the protein expression level of HO-1, C-caspase 3, total PARP, and cleaved PARP were measured by Western blotting, and β-actin was used as loading control ( A ). ( B ) Quantitative analysis of the blots in ( A ). ( C ) The apoptotic rates of cells were determined by Annexin V-PI staining and flow cytometry for quantification. ( D ) HO activity was measured in ST88-14 and S462 cells at 48 hours treatment of 40.0 μmol L −1 flaccidoside II with/without the performance of ERK-1/2 inhibitors PD98059 or p38 inhibitor SB203580, respectively. Each bar represents the mean ± SD of at least four independent experiments. * P

    Article Snippet: The following antibodies were used: anti-HO-1 (Epitomics, Burlingame, CA, USA), antiphospho-p38 (Cell Signaling Technology, Beverly, MA, USA), antiphospho-ERK-1/2 (Cell Signaling Technology), antiphospho-JNK (Santa Cruz Biotechnology Inc., Dallas, TX, USA), anti-p38 (Cell Signaling Technology), anti-ERK-1/2 (Cell Signaling Technology), anti-JNK (Santa Cruz Biotechnology Inc.), and anti-GAPDH (Abcam, Cambridge, MA, USA).

    Techniques: Expressing, Western Blot, Staining, Flow Cytometry, Cytometry, Activity Assay

    Effect of SPD-induced HO-1 inhibition on cell death. Cells were incubated in the absence or presence of ZnPP, siRNA of HO-1 or Nrf2 for 24 h before the indicated tests were performed. SPD-stimulated HUVECs were pretreated with or without ZnPP, HO-1 or Nrf2 siRNA. Protective effect of HO-1 induction on cell death was determined by in situ terminal nick-end labeling (TUNEL). Representative images illustrate fluorescent TUNEL (green) staining of cells cultured for 24 h before the indicated tests were performed. The percentage of TUNEL-positive cells per total cell count in each sample was calculated. At least 100 cells from three random fields were counted in each experiment. Data are the mean ± SD of triplicate experiments. * P

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Hemeoxygenase-1 Mediates an Adaptive Response to Spermidine-Induced Cell Death in Human Endothelial Cells

    doi: 10.1155/2013/238734

    Figure Lengend Snippet: Effect of SPD-induced HO-1 inhibition on cell death. Cells were incubated in the absence or presence of ZnPP, siRNA of HO-1 or Nrf2 for 24 h before the indicated tests were performed. SPD-stimulated HUVECs were pretreated with or without ZnPP, HO-1 or Nrf2 siRNA. Protective effect of HO-1 induction on cell death was determined by in situ terminal nick-end labeling (TUNEL). Representative images illustrate fluorescent TUNEL (green) staining of cells cultured for 24 h before the indicated tests were performed. The percentage of TUNEL-positive cells per total cell count in each sample was calculated. At least 100 cells from three random fields were counted in each experiment. Data are the mean ± SD of triplicate experiments. * P

    Article Snippet: Anti-HO-1 antibody was obtained from Epitomics (Burlingame, CA, USA).

    Techniques: Inhibition, Incubation, In Situ, End Labeling, TUNEL Assay, Staining, Cell Culture, Cell Counting

    A scheme showing a proposed cytoprotective mechanism against SPD-induced oxidative stress by HO-1 induction. During the degradation of spermidine (SPD) to putrescine by serum amine oxidase, aldehydes, H 2 O 2 , and NH 3 are produced, which induce oxidative stress. It triggers Nrf2 translocation and ARE binding through PI3K/Akt signaling. Nrf2 activation upregulates HO-1 expression. HO-1 catalyzed heme to biliverdin and bilirubin, with the concurrent release of bioactive molecules, such as ferritin and CO. Heme metabolites exert adaptive response and protect cells against SPD-induced oxidative stress.

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Hemeoxygenase-1 Mediates an Adaptive Response to Spermidine-Induced Cell Death in Human Endothelial Cells

    doi: 10.1155/2013/238734

    Figure Lengend Snippet: A scheme showing a proposed cytoprotective mechanism against SPD-induced oxidative stress by HO-1 induction. During the degradation of spermidine (SPD) to putrescine by serum amine oxidase, aldehydes, H 2 O 2 , and NH 3 are produced, which induce oxidative stress. It triggers Nrf2 translocation and ARE binding through PI3K/Akt signaling. Nrf2 activation upregulates HO-1 expression. HO-1 catalyzed heme to biliverdin and bilirubin, with the concurrent release of bioactive molecules, such as ferritin and CO. Heme metabolites exert adaptive response and protect cells against SPD-induced oxidative stress.

    Article Snippet: Anti-HO-1 antibody was obtained from Epitomics (Burlingame, CA, USA).

    Techniques: Produced, Translocation Assay, Binding Assay, Activation Assay, Expressing

    Induction of HO-1 by SPD in HUVECs. After treatment of HUVECs with various concentrations of SPD (a) at various time intervals (b), cell lysates were prepared, and 20 μ g samples of proteins were subjected to Western blotting using the anti-HO-1 antibody, and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody as a loading control. Representative data from three independent experiments are shown. After treatment of HUVECs with various concentrations of SPD (c) at various time intervals (d), cell lysates were prepared. The total RNA was extracted and analyzed by RT-PCR. The amplified RT-PCR product was visualized on 1% agarose gel. Representative data from three independent experiments are shown.

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Hemeoxygenase-1 Mediates an Adaptive Response to Spermidine-Induced Cell Death in Human Endothelial Cells

    doi: 10.1155/2013/238734

    Figure Lengend Snippet: Induction of HO-1 by SPD in HUVECs. After treatment of HUVECs with various concentrations of SPD (a) at various time intervals (b), cell lysates were prepared, and 20 μ g samples of proteins were subjected to Western blotting using the anti-HO-1 antibody, and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody as a loading control. Representative data from three independent experiments are shown. After treatment of HUVECs with various concentrations of SPD (c) at various time intervals (d), cell lysates were prepared. The total RNA was extracted and analyzed by RT-PCR. The amplified RT-PCR product was visualized on 1% agarose gel. Representative data from three independent experiments are shown.

    Article Snippet: Anti-HO-1 antibody was obtained from Epitomics (Burlingame, CA, USA).

    Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis

    Blockage of SPD-induced HO-1 protein expression by aminoguanidine, NAC, or PI3K/Akt inhibitor. Cells were pre-treated with aminoguanidine (a), NAC (b), or LY 294002 (PI3K/Akt inhibitor) (c) 1 h prior to the treatment of SPD. After 24 h incubation, cell lysates were prepared, and 20 μ g samples of proteins were subjected to Western blotting, using anti-HO-1 antibody and anti-GAPDH.

    Journal: Oxidative Medicine and Cellular Longevity

    Article Title: Hemeoxygenase-1 Mediates an Adaptive Response to Spermidine-Induced Cell Death in Human Endothelial Cells

    doi: 10.1155/2013/238734

    Figure Lengend Snippet: Blockage of SPD-induced HO-1 protein expression by aminoguanidine, NAC, or PI3K/Akt inhibitor. Cells were pre-treated with aminoguanidine (a), NAC (b), or LY 294002 (PI3K/Akt inhibitor) (c) 1 h prior to the treatment of SPD. After 24 h incubation, cell lysates were prepared, and 20 μ g samples of proteins were subjected to Western blotting, using anti-HO-1 antibody and anti-GAPDH.

    Article Snippet: Anti-HO-1 antibody was obtained from Epitomics (Burlingame, CA, USA).

    Techniques: Expressing, Incubation, Western Blot

    KMUP-1 treatment elevated the expression of HO-1 protein. ( A ) H9c2 cells were preincubated with ZnPP at 10 μM for 1 h, followed by co-treatment with KMUP-1 or ( B ) combination with 100 nM of ET-1. Western blot analysis was conducted to measure the induction of HO-1 protein; ( C ) The expression of calcineurin A was analyzed. # p

    Journal: Molecules

    Article Title: KMUP-1 Attenuates Endothelin-1-Induced Cardiomyocyte Hypertrophy through Activation of Heme Oxygenase-1 and Suppression of the Akt/GSK-3β, Calcineurin/NFATc4 and RhoA/ROCK Pathways

    doi: 10.3390/molecules200610435

    Figure Lengend Snippet: KMUP-1 treatment elevated the expression of HO-1 protein. ( A ) H9c2 cells were preincubated with ZnPP at 10 μM for 1 h, followed by co-treatment with KMUP-1 or ( B ) combination with 100 nM of ET-1. Western blot analysis was conducted to measure the induction of HO-1 protein; ( C ) The expression of calcineurin A was analyzed. # p

    Article Snippet: Anti-HO-1, anti-GSK3β (Ser9 ), anti-calcineurin and anti-ROCK-II were purchased from BD Biosciences (San Diego, CA, USA).

    Techniques: Expressing, Western Blot

    Proposed signaling pathways involved in the protective effects of KMUP-1 on ET-1-induced cardiomyocyte hypertrophy. The underlying mechanisms are associated with inhibition of p-ERK1/2, p-Akt, p-GSK-3β, calcineurin A, nuclear NFATc4 expression and Rho A translocation, subsequent AP-1 expression and up-regulation of HO-1.

    Journal: Molecules

    Article Title: KMUP-1 Attenuates Endothelin-1-Induced Cardiomyocyte Hypertrophy through Activation of Heme Oxygenase-1 and Suppression of the Akt/GSK-3β, Calcineurin/NFATc4 and RhoA/ROCK Pathways

    doi: 10.3390/molecules200610435

    Figure Lengend Snippet: Proposed signaling pathways involved in the protective effects of KMUP-1 on ET-1-induced cardiomyocyte hypertrophy. The underlying mechanisms are associated with inhibition of p-ERK1/2, p-Akt, p-GSK-3β, calcineurin A, nuclear NFATc4 expression and Rho A translocation, subsequent AP-1 expression and up-regulation of HO-1.

    Article Snippet: Anti-HO-1, anti-GSK3β (Ser9 ), anti-calcineurin and anti-ROCK-II were purchased from BD Biosciences (San Diego, CA, USA).

    Techniques: Inhibition, Expressing, Translocation Assay

    a Quantification of densitometric analysis of immunohistochemistry. Data are presented as median (IQR). Representative pictures ×10. b CSE YGP, c CSE FBM sham, d CSE FBM sepsis, e HO-1 YGP, f HO-1 FBM sham, and ( g ) HO-1 FBM sepsis. b – g show a cross-section of the vessel wall with the luminal side to the right . h eNOS YGP, i eNOS FBM sham, and ( j ) eNOS FBM sepsis. h – j show a cross-section of the vessel wall with the luminal side to the upper left corner

    Journal: Intensive Care Medicine Experimental

    Article Title: Cardiovascular disease and resuscitated septic shock lead to the downregulation of the H2S-producing enzyme cystathionine-γ-lyase in the porcine coronary artery

    doi: 10.1186/s40635-017-0131-8

    Figure Lengend Snippet: a Quantification of densitometric analysis of immunohistochemistry. Data are presented as median (IQR). Representative pictures ×10. b CSE YGP, c CSE FBM sham, d CSE FBM sepsis, e HO-1 YGP, f HO-1 FBM sham, and ( g ) HO-1 FBM sepsis. b – g show a cross-section of the vessel wall with the luminal side to the right . h eNOS YGP, i eNOS FBM sham, and ( j ) eNOS FBM sepsis. h – j show a cross-section of the vessel wall with the luminal side to the upper left corner

    Article Snippet: After heat-induced antigen retrieval, the slides were blocked with 10% normal sera (Jackson ImmunoResearch) before incubating in primary antibody (1° ab, anti-nitrotyrosine (Millipore)) and CSE: anti-CTH (Abnova), anti-CBS (Santa Cruz Biotechnology), anti-MPST (Sigma), anti-adipophilin (Progen), anti-eNOS (BD), and anti-HO-1 (Abnova).

    Techniques: Immunohistochemistry

    DMF  increases expression of antioxidant enzymes in  RAW  264.7 cells. Gene expression for  NQO 1  ( A ),  HO ‐1  ( B ) and  GCS  ( C ) are shown. * P 

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Dimethyl fumarate inhibits osteoclasts via attenuation of reactive oxygen species signalling by augmented antioxidation

    doi: 10.1111/jcmm.13367

    Figure Lengend Snippet: DMF increases expression of antioxidant enzymes in RAW 264.7 cells. Gene expression for NQO 1 ( A ), HO ‐1 ( B ) and GCS ( C ) are shown. * P 

    Article Snippet: The primary antibodies for these experiments were anti‐Nrf2 (1/1000 dilution; Santa Cruz Biotechnology Inc.), anti‐histone H3 (1/4000; Cell Signaling Technology Japan, Tokyo, Japan), anti HO‐1 (1/5000; StressMarq Biosciences Inc., Victoria, BC, Canada), anti‐NQO1 (1/5000; Abcam plc, Cambridge, MA, USA) and anti‐GCS (1/5000; Thermo Fisher Scientific Inc.).

    Techniques: Expressing

    Expression of cytoprotective genes was up-regulated by Nrf2 overexpression and down-regulated by Keap1 overexpression or Nrf2 knockdown. The expressions of HO-1, NQO1, and GCS were examined by real time PCR at day 2 ( A ). Gene expression was calibrated using the Rps18 housekeeping gene, and values indicating the fold-change from control are shown. The data represent the means of three independent experiments performed in triplicate. *, p

    Journal: The Journal of Biological Chemistry

    Article Title: The Keap1/Nrf2 Protein Axis Plays a Role in Osteoclast Differentiation by Regulating Intracellular Reactive Oxygen Species Signaling *

    doi: 10.1074/jbc.M113.478545

    Figure Lengend Snippet: Expression of cytoprotective genes was up-regulated by Nrf2 overexpression and down-regulated by Keap1 overexpression or Nrf2 knockdown. The expressions of HO-1, NQO1, and GCS were examined by real time PCR at day 2 ( A ). Gene expression was calibrated using the Rps18 housekeeping gene, and values indicating the fold-change from control are shown. The data represent the means of three independent experiments performed in triplicate. *, p

    Article Snippet: The antibodies used in these experiments were anti HO-1 antibody (StressMarq Biosciences Inc., Victoria, British Columbia, Canada), anti-NQO1 antibody (Abcam plc, Cambridge, MA), and anti-GCS antibody (Thermo Fisher Scientific Inc.).

    Techniques: Expressing, Over Expression, Real-time Polymerase Chain Reaction

    Cytoprotective enzymes are down-regulated by RANKL stimulation. The expressions of HO-1, NQO1, and GCS were examined by real time PCR at day 1 ( A ) and day 2 ( B ). Gene expression was calibrated using the RPS18 housekeeping gene, and values indicating the fold-change from control are shown. Open bars represent controls, and closed bars indicate stimulation by RANKL. The data shown are representative of three independent experiments performed in triplicate. *, p

    Journal: The Journal of Biological Chemistry

    Article Title: The Keap1/Nrf2 Protein Axis Plays a Role in Osteoclast Differentiation by Regulating Intracellular Reactive Oxygen Species Signaling *

    doi: 10.1074/jbc.M113.478545

    Figure Lengend Snippet: Cytoprotective enzymes are down-regulated by RANKL stimulation. The expressions of HO-1, NQO1, and GCS were examined by real time PCR at day 1 ( A ) and day 2 ( B ). Gene expression was calibrated using the RPS18 housekeeping gene, and values indicating the fold-change from control are shown. Open bars represent controls, and closed bars indicate stimulation by RANKL. The data shown are representative of three independent experiments performed in triplicate. *, p

    Article Snippet: The antibodies used in these experiments were anti HO-1 antibody (StressMarq Biosciences Inc., Victoria, British Columbia, Canada), anti-NQO1 antibody (Abcam plc, Cambridge, MA), and anti-GCS antibody (Thermo Fisher Scientific Inc.).

    Techniques: Real-time Polymerase Chain Reaction, Expressing

    Intracellular heme oxygenase 1 (HO-1) in primary human tracheo-bronchial epithelial cells 24 h after treatment with sulfur mustard (SM) vapor. HO-1 levels in the medium were measured by western blotting, and normalized to β-actin levels in the same blots following stripping and re-probing. Incubator control (inc ctl): cells maintained in the CO 2 incubator during exposure of the remaining cultures in a glove box. 0: Cells exposed to the vapor from 10 μl of ethanol. SM concentrations are the estimated maximal concentration possible in the vapor phase as the result of the amount present in 10 μl of the dilution applied to the filter.

    Journal: Inhalation toxicology

    Article Title: Sulfur mustard vapor effects on differentiated human lung cells

    doi: 10.3109/08958378.2010.493901

    Figure Lengend Snippet: Intracellular heme oxygenase 1 (HO-1) in primary human tracheo-bronchial epithelial cells 24 h after treatment with sulfur mustard (SM) vapor. HO-1 levels in the medium were measured by western blotting, and normalized to β-actin levels in the same blots following stripping and re-probing. Incubator control (inc ctl): cells maintained in the CO 2 incubator during exposure of the remaining cultures in a glove box. 0: Cells exposed to the vapor from 10 μl of ethanol. SM concentrations are the estimated maximal concentration possible in the vapor phase as the result of the amount present in 10 μl of the dilution applied to the filter.

    Article Snippet: Antigen was detected using 1:1000 of anti-HO-1 (Stressgen™, Assay Designs, Ann Arbor, Michigan) and horse-radish peroxidase-conjugated (HRP-conjugated) anti-rabbit secondary antibody (Sigma Chemical Co., St. Louis, MO).

    Techniques: Western Blot, Stripping Membranes, CTL Assay, Concentration Assay

    Summary of paraquat redox cycling and detoxification of ROI by cellular antioxidants Upper panel : Paraquat redox cycling reactions. Paraquat undergoes a one electron reduction to the paraquat radical via the oxidation of NAD(P)H to NAD(P) + by NAD(P)H oxidase (Reaction 1). The paraquat radical is then immediately oxidized to the parent compound with the transfer of an electron to molecular oxygen, forming superoxide anion (Reaction 2). Lower panel : Detoxification of ROI by enzymatic antioxidants. Superoxide anion is metabolized to hydrogen peroxide by SOD. Hydrogen peroxide is detoxified by catalase and/or various peroxidases including GPx-1. Hydrogen peroxide has been implicated in the mobilization of heme, an oxidizing agent, which is then degraded by heme oxygenases including HO-1 to biliverdin, ferric iron and carbon monoxide. Biliverdin is further metabolized to bilirubin and ferric iron is removed through sequestration by ferritin. In the presence of transition metals such as iron or copper, hydrogen peroxide can also be converted to hydroxyl radicals. The zinc-mediated free radical scavenging function of MT-2 serves as a mechanism for removal of hydroxyl radicals. Oxidized proteins and lipids can be conjugated with glutathione by the GST enzymes to facilitate cellular elimination.

    Journal: Toxicology and applied pharmacology

    Article Title: Increased oxidative stress and antioxidant expression in mouse keratinocytes following exposure to paraquat

    doi: 10.1016/j.taap.2008.05.014

    Figure Lengend Snippet: Summary of paraquat redox cycling and detoxification of ROI by cellular antioxidants Upper panel : Paraquat redox cycling reactions. Paraquat undergoes a one electron reduction to the paraquat radical via the oxidation of NAD(P)H to NAD(P) + by NAD(P)H oxidase (Reaction 1). The paraquat radical is then immediately oxidized to the parent compound with the transfer of an electron to molecular oxygen, forming superoxide anion (Reaction 2). Lower panel : Detoxification of ROI by enzymatic antioxidants. Superoxide anion is metabolized to hydrogen peroxide by SOD. Hydrogen peroxide is detoxified by catalase and/or various peroxidases including GPx-1. Hydrogen peroxide has been implicated in the mobilization of heme, an oxidizing agent, which is then degraded by heme oxygenases including HO-1 to biliverdin, ferric iron and carbon monoxide. Biliverdin is further metabolized to bilirubin and ferric iron is removed through sequestration by ferritin. In the presence of transition metals such as iron or copper, hydrogen peroxide can also be converted to hydroxyl radicals. The zinc-mediated free radical scavenging function of MT-2 serves as a mechanism for removal of hydroxyl radicals. Oxidized proteins and lipids can be conjugated with glutathione by the GST enzymes to facilitate cellular elimination.

    Article Snippet: Anti- HO-1 antibodies were purchased from Assay Designs (Ann Arbor, MI), anti-catalase antibodies were from Abcam (Cambridge, MA) and Cu,Zn-SOD antibodies from Santa Cruz Biotechnology (Santa Cruz, CA).

    Techniques:

    SFN pretreatment and cadmium-induced Nrf2/ARE (antioxidant response element) signaling in mouse testes. ( A ) Nuclear factor-erythroid 2-related factor 2 (Nrf2); ( B ) NAD(P)H:quinone oxidoreductase 1 (NQO1); ( C ) hemeoxygenase-1 (HO-1); ( D ) γ-glutamyl cysteine synthetase (γ-GCS); and ( E ) glutathione peroxidase (GSH-Px). Values represent mean ± SEM in each group of 10 mice; * p

    Journal: International Journal of Molecular Sciences

    Article Title: Sulforaphane Prevents Testicular Damage in Kunming Mice Exposed to Cadmium via Activation of Nrf2/ARE Signaling Pathways

    doi: 10.3390/ijms17101703

    Figure Lengend Snippet: SFN pretreatment and cadmium-induced Nrf2/ARE (antioxidant response element) signaling in mouse testes. ( A ) Nuclear factor-erythroid 2-related factor 2 (Nrf2); ( B ) NAD(P)H:quinone oxidoreductase 1 (NQO1); ( C ) hemeoxygenase-1 (HO-1); ( D ) γ-glutamyl cysteine synthetase (γ-GCS); and ( E ) glutathione peroxidase (GSH-Px). Values represent mean ± SEM in each group of 10 mice; * p

    Article Snippet: Total protein (50 µg) was fractionated on 10% SDS-PAGE gels, transferred electrophoretically onto a polyvinylidene difluoride membrane (Bio-Rad, Hercules, CA, USA), and blocked with 5% nonfat dry milk in TBST buffer for 2 h. Membranes were incubated overnight at 4 °C with anti-Nrf2 (1:500, Santa), anti-GSH-PX (1:500, Santa), anti-γ-GCS (1:500, Santa), anti-HO-1 (1:300, Sangon Biotech), and anti-NQO1 (1:1000, Sangon Biotech) antibodies.

    Techniques: Mouse Assay

    DMF suppresses LPS-induced reactive MG. Primary MG were stimulated by LPS (100 ng/ml) in the absence or presence of DMF for various time periods. a Microglial surface markers, CD86, CD80, and CD40, were examined at 18 h after stimulation using FACS analysis. Representative flow plots from five independent experiments are shown. ISO, the isotype control antibody. MED, cell culture medium. b Morphological changes of MG at 18 h after stimulation were examined using Iba1 immunostaining. Arrows indicate activated MG with increased expression of Iba1 and enlarged cell bodies. Cell nuclei were stained blue with Hoechst 33342. Scale bar, 20 μm. c The expressions of DMF-induced Nrf2 target genes were examined at 3 h after stimulation. The mRNA levels of hmox1 , nqo1 , gclc , and gclm were quantified using qPCR. Data presented are from 3 to 5 independent experiments. * p

    Journal: Journal of Neuroinflammation

    Article Title: Dimethyl fumarate attenuates reactive microglia and long-term memory deficits following systemic immune challenge

    doi: 10.1186/s12974-018-1125-5

    Figure Lengend Snippet: DMF suppresses LPS-induced reactive MG. Primary MG were stimulated by LPS (100 ng/ml) in the absence or presence of DMF for various time periods. a Microglial surface markers, CD86, CD80, and CD40, were examined at 18 h after stimulation using FACS analysis. Representative flow plots from five independent experiments are shown. ISO, the isotype control antibody. MED, cell culture medium. b Morphological changes of MG at 18 h after stimulation were examined using Iba1 immunostaining. Arrows indicate activated MG with increased expression of Iba1 and enlarged cell bodies. Cell nuclei were stained blue with Hoechst 33342. Scale bar, 20 μm. c The expressions of DMF-induced Nrf2 target genes were examined at 3 h after stimulation. The mRNA levels of hmox1 , nqo1 , gclc , and gclm were quantified using qPCR. Data presented are from 3 to 5 independent experiments. * p

    Article Snippet: For the western blot analysis, rabbit anti-Nrf2 and anti-HO-1 antibodies were purchased from Proteintech (Chicago, IL).

    Techniques: FACS, Flow Cytometry, Cell Culture, Immunostaining, Expressing, Staining, Real-time Polymerase Chain Reaction