glyceraldehyde 3 phosphate dehydrogenase gapdh  (Qiagen)

 
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    Structured Review

    Qiagen glyceraldehyde 3 phosphate dehydrogenase gapdh
    HCV NS5A enhanced MG132-induced NF-κB p65 nuclear translocation. A , HepG2 control and HepG2-NS5Acells were cultured for 6 hours with or without 5 μM MG132. Confocal microscope high-power view demonstrates that NF-κB p65 nuclear localization was detected (x200). Nuclear staining was performed with Hoechst 33342, trihydrochloride, trihydrate (blue). Localization of NF-κB p65 was detected with anti-NF-κB p65 primary antibody, and Alexa-Fluor-555 secondary antibody (red). Merge images of A and B were superimposed digitally to allow for fine comparisons. NF-κB p65 nuclear translocation was observed (pink). B , The number of NF-κB p65 nuclear-translocated cells per field was counted at low-power view (40X). C , Huh7 cells infected with or without HCV JFH1 genotype 2 strain were incubated for 6 hours with or without 5 μM MG132. Nuclear staining was performed with Hoechst 33342, trihydrochloride, trihydrate (blue). NF-κB p65 nuclear localization was detected with anti-NF-κB p65 primary antibody and anti-rabbit Alexa-Fluor-555 secondary antibody (red). HCV was detected using anti-HCV core primary antibody and an Alexa-Fluor-488 anti-mouse secondary antibody (green). D , HCV NS5A expression enhances phosphorylation of IκBα. Western blot analyses of phosphorylated IκBα (Ser32) (P-IκBα), IκBα and <t>glyceraldehyde-3-phosphate</t> dehydrogenase <t>(GAPDH)</t> expression in HepG2 control or HepG2-NS5A cells at 8 hours after treatment with or without MG132.
    Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh, supplied by Qiagen, used in various techniques. Bioz Stars score: 93/100, based on 64 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Hepatitis C Virus Nonstructural Protein 5A Inhibits MG132-Induced Apoptosis of Hepatocytes in Line with NF-κB-Nuclear Translocation"

    Article Title: Hepatitis C Virus Nonstructural Protein 5A Inhibits MG132-Induced Apoptosis of Hepatocytes in Line with NF-κB-Nuclear Translocation

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0131973

    HCV NS5A enhanced MG132-induced NF-κB p65 nuclear translocation. A , HepG2 control and HepG2-NS5Acells were cultured for 6 hours with or without 5 μM MG132. Confocal microscope high-power view demonstrates that NF-κB p65 nuclear localization was detected (x200). Nuclear staining was performed with Hoechst 33342, trihydrochloride, trihydrate (blue). Localization of NF-κB p65 was detected with anti-NF-κB p65 primary antibody, and Alexa-Fluor-555 secondary antibody (red). Merge images of A and B were superimposed digitally to allow for fine comparisons. NF-κB p65 nuclear translocation was observed (pink). B , The number of NF-κB p65 nuclear-translocated cells per field was counted at low-power view (40X). C , Huh7 cells infected with or without HCV JFH1 genotype 2 strain were incubated for 6 hours with or without 5 μM MG132. Nuclear staining was performed with Hoechst 33342, trihydrochloride, trihydrate (blue). NF-κB p65 nuclear localization was detected with anti-NF-κB p65 primary antibody and anti-rabbit Alexa-Fluor-555 secondary antibody (red). HCV was detected using anti-HCV core primary antibody and an Alexa-Fluor-488 anti-mouse secondary antibody (green). D , HCV NS5A expression enhances phosphorylation of IκBα. Western blot analyses of phosphorylated IκBα (Ser32) (P-IκBα), IκBα and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression in HepG2 control or HepG2-NS5A cells at 8 hours after treatment with or without MG132.
    Figure Legend Snippet: HCV NS5A enhanced MG132-induced NF-κB p65 nuclear translocation. A , HepG2 control and HepG2-NS5Acells were cultured for 6 hours with or without 5 μM MG132. Confocal microscope high-power view demonstrates that NF-κB p65 nuclear localization was detected (x200). Nuclear staining was performed with Hoechst 33342, trihydrochloride, trihydrate (blue). Localization of NF-κB p65 was detected with anti-NF-κB p65 primary antibody, and Alexa-Fluor-555 secondary antibody (red). Merge images of A and B were superimposed digitally to allow for fine comparisons. NF-κB p65 nuclear translocation was observed (pink). B , The number of NF-κB p65 nuclear-translocated cells per field was counted at low-power view (40X). C , Huh7 cells infected with or without HCV JFH1 genotype 2 strain were incubated for 6 hours with or without 5 μM MG132. Nuclear staining was performed with Hoechst 33342, trihydrochloride, trihydrate (blue). NF-κB p65 nuclear localization was detected with anti-NF-κB p65 primary antibody and anti-rabbit Alexa-Fluor-555 secondary antibody (red). HCV was detected using anti-HCV core primary antibody and an Alexa-Fluor-488 anti-mouse secondary antibody (green). D , HCV NS5A expression enhances phosphorylation of IκBα. Western blot analyses of phosphorylated IκBα (Ser32) (P-IκBα), IκBα and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression in HepG2 control or HepG2-NS5A cells at 8 hours after treatment with or without MG132.

    Techniques Used: Translocation Assay, Cell Culture, Microscopy, Staining, Infection, Incubation, Expressing, Western Blot

    Related Articles

    SYBR Green Assay:

    Article Title: Promotion of Functional Nerve Regeneration by Inhibition of Microtubule Detyrosination
    Article Snippet: .. NFκB inhibitor α (IκBα) and glyceraldehyde 3-phosphate dehydrogenase (gapdh) were amplified using respective QuantiTect primers (QIAGEN) and SYBR Green PCR Master Mix (Applied Biosystems) on an Applied Biosystems 7500 real-time PCR system (Thermo Fisher Scientific) using 45 amplification cycles. .. PCR specificity was verified with the dissociation curve analysis feature.

    Multiplex Assay:

    Article Title: Suberoyl bis-hydroxamic acid reactivates Kaposi’s sarcoma-associated herpesvirus through histone acetylation and induces apoptosis in lymphoma cells
    Article Snippet: .. RNA samples were subjected to TaqMan-based real-time RT-PCR analysis to detect the mRNA of RTA, vIL-6 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) using QuantiTect Multiplex RT-PCR Kits (Qiagen) as described previously ( , ). .. Western blot analysis Protein extraction and immunoblotting were performed as described previously, with some modifications ( ).

    Article Title: Protection Provided by a Herpes Simplex Virus 2 (HSV-2) Glycoprotein C and D Subunit Antigen Vaccine against Genital HSV-2 Infection in HSV-1-Seropositive Guinea Pigs
    Article Snippet: .. DNA samples and oligonucleotide primer-probe sets specific for gG2 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were mixed with Quantitect Multiplex PCR no-Rox master mix (Qiagen). .. Reactions were carried out in a Stratagene Mx3005P real-time PCR system and analyzed with Stratagene MxPro software.

    Amplification:

    Article Title: Promotion of Functional Nerve Regeneration by Inhibition of Microtubule Detyrosination
    Article Snippet: .. NFκB inhibitor α (IκBα) and glyceraldehyde 3-phosphate dehydrogenase (gapdh) were amplified using respective QuantiTect primers (QIAGEN) and SYBR Green PCR Master Mix (Applied Biosystems) on an Applied Biosystems 7500 real-time PCR system (Thermo Fisher Scientific) using 45 amplification cycles. .. PCR specificity was verified with the dissociation curve analysis feature.

    Marker:

    Article Title: Transition of responsive mechanosensitive elements from focal adhesions to adherens junctions on epithelial differentiation
    Article Snippet: .. Taq Man assays against α-catenin, paxillin, talin, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as constitutively expressed marker (Qiagen, Hilden, Germany) were used. .. For reincorporation of human full-length α-catenin (h-αCat) and ΔvinBD α-catenin, cells were knocked down for endogenous α-catenin first by using a mouse-specific siRNA (siPOOL5; SiTools biotech, Martinsried, Germany).

    Quantitative RT-PCR:

    Article Title: Suberoyl bis-hydroxamic acid reactivates Kaposi’s sarcoma-associated herpesvirus through histone acetylation and induces apoptosis in lymphoma cells
    Article Snippet: .. RNA samples were subjected to TaqMan-based real-time RT-PCR analysis to detect the mRNA of RTA, vIL-6 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) using QuantiTect Multiplex RT-PCR Kits (Qiagen) as described previously ( , ). .. Western blot analysis Protein extraction and immunoblotting were performed as described previously, with some modifications ( ).

    Real-time Polymerase Chain Reaction:

    Article Title: Promotion of Functional Nerve Regeneration by Inhibition of Microtubule Detyrosination
    Article Snippet: .. NFκB inhibitor α (IκBα) and glyceraldehyde 3-phosphate dehydrogenase (gapdh) were amplified using respective QuantiTect primers (QIAGEN) and SYBR Green PCR Master Mix (Applied Biosystems) on an Applied Biosystems 7500 real-time PCR system (Thermo Fisher Scientific) using 45 amplification cycles. .. PCR specificity was verified with the dissociation curve analysis feature.

    Article Title: Simultaneous Amelioratation of Colitis and Liver Injury in Mice by Bifidobacterium longum LC67 and Lactobacillus plantarum LC27
    Article Snippet: .. Real time PCR for IL-10, IL-17, Foxp3, RAR-related orphan receptor gamma t (RORγt), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was performed as described previously , utilizing Qiagen thermal cycler, which used SYBER premix agents. .. Thermal cycling conditions were as follows: activation of DNA polymerase at 95 °C for 5 min, followed by 36 cycles of denaturation and amplification at 95 °C for 5 s and 63 °C for 30 s, respectively.

    Polymerase Chain Reaction:

    Article Title: Promotion of Functional Nerve Regeneration by Inhibition of Microtubule Detyrosination
    Article Snippet: .. NFκB inhibitor α (IκBα) and glyceraldehyde 3-phosphate dehydrogenase (gapdh) were amplified using respective QuantiTect primers (QIAGEN) and SYBR Green PCR Master Mix (Applied Biosystems) on an Applied Biosystems 7500 real-time PCR system (Thermo Fisher Scientific) using 45 amplification cycles. .. PCR specificity was verified with the dissociation curve analysis feature.

    Article Title: Hepatitis C Virus Nonstructural Protein 5A Inhibits MG132-Induced Apoptosis of Hepatocytes in Line with NF-κB-Nuclear Translocation
    Article Snippet: .. PCR was performed on cDNA templates using primers specific for B-cell CLL/lymphoma (BCL2), BCL2-like 1 (BCL2L1/BCL-X/BCLXL) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), which were purchased from Qiagen. .. A Human NF-κB Signaling Targets Real-time RT-PCR Array was performed according to the manufacturer's protocol.

    Article Title: Protection Provided by a Herpes Simplex Virus 2 (HSV-2) Glycoprotein C and D Subunit Antigen Vaccine against Genital HSV-2 Infection in HSV-1-Seropositive Guinea Pigs
    Article Snippet: .. DNA samples and oligonucleotide primer-probe sets specific for gG2 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) were mixed with Quantitect Multiplex PCR no-Rox master mix (Qiagen). .. Reactions were carried out in a Stratagene Mx3005P real-time PCR system and analyzed with Stratagene MxPro software.

    Expressing:

    Article Title: Prostate-derived Ets transcription factor (PDEF) downregulates survivin expression and inhibits breast cancer cell growth in vitro and xenograft tumor formation in vivo
    Article Snippet: .. The mRNA expression for PDEF, survivin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was determined using a one-step RT PCR kit (Qiagen) as previously described [ ]. .. The primers used for RT-PCR were: 5′-ATG GGC AGC GCC AGC CCG GGT C-3′(forward) and 5′-TCA GAT GGG GTG CAC GAA CTG GT-3′(reverse) for PDEF PCR products (1008 bp), 5′-GAG GCT GGC TTC ATC CAC TG-3′(forward) and 5′-CAG CTG CTC GAT GGC ACG GC-3′(reverse) for survivin PCR products (299 bp), and 5′-GCT TCC CGT TCT CAG CCT TGA C-3′(forward) and 5′-ATG GGA AGG TGA AGG TCG GAG-3′(reverse) for GAPDH PCR products (195 bp, internal control). )

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Prostate-derived Ets transcription factor (PDEF) downregulates survivin expression and inhibits breast cancer cell growth in vitro and xenograft tumor formation in vivo
    Article Snippet: .. The mRNA expression for PDEF, survivin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was determined using a one-step RT PCR kit (Qiagen) as previously described [ ]. .. The primers used for RT-PCR were: 5′-ATG GGC AGC GCC AGC CCG GGT C-3′(forward) and 5′-TCA GAT GGG GTG CAC GAA CTG GT-3′(reverse) for PDEF PCR products (1008 bp), 5′-GAG GCT GGC TTC ATC CAC TG-3′(forward) and 5′-CAG CTG CTC GAT GGC ACG GC-3′(reverse) for survivin PCR products (299 bp), and 5′-GCT TCC CGT TCT CAG CCT TGA C-3′(forward) and 5′-ATG GGA AGG TGA AGG TCG GAG-3′(reverse) for GAPDH PCR products (195 bp, internal control). )

    Article Title: Suberoyl bis-hydroxamic acid reactivates Kaposi’s sarcoma-associated herpesvirus through histone acetylation and induces apoptosis in lymphoma cells
    Article Snippet: .. RNA samples were subjected to TaqMan-based real-time RT-PCR analysis to detect the mRNA of RTA, vIL-6 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) using QuantiTect Multiplex RT-PCR Kits (Qiagen) as described previously ( , ). .. Western blot analysis Protein extraction and immunoblotting were performed as described previously, with some modifications ( ).

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    Qiagen glyceraldehyde 3 phosphate dehydrogenase
    Hp reduces Hb-induced cellular activation, inflammatory response and oxidative stress in the choroid plexus. Rabbit pups with confirmed IVH, injected intraventricularly with Hp (grey bars; n = 6) or sham (white bars; n = 6) 6 hours after confirmation of the bleeding, were euthanized at 24 hours of age and the brains were removed from the skulls and the choroid plexus was carefully removed from the lateral ventricles, snap frozen, and the mRNA expression of IL1R1 (A) , FAS (B) , NF-Κβ (C) , MCP-1 (D) , IL-8 (E) , IL-1β (F) , TNFα (G) , IL-6 (H) and HO-1 (I) were subsequently analyzed with real-time PCR, as described in Materials and Methods. mRNA expression for respective gene was normalized against those of <t>glyceraldehyde-3-phosphate</t> dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against samples from control pups. Results are presented as box plots displaying medians and 25 th and 75 th percentiles. Differences between IVH + Hp versus IVH + Sham at 24 hours were analyzed using the Mann-Whitney U -test. * P
    Glyceraldehyde 3 Phosphate Dehydrogenase, supplied by Qiagen, used in various techniques. Bioz Stars score: 93/100, based on 67 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/glyceraldehyde 3 phosphate dehydrogenase/product/Qiagen
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    92
    Qiagen human glyceraldehyde 3 phosphate dehydrogenase
    BAY 11-7082 reduced the acidic bile-induced gene expression profiling of NF-κB signaling pathway in treated normal human hypopharyngeal cells Heat maps were obtained by RT 2 -Profiler PCR array analysis for NF-κB signaling. The genes were clustered based on their biological role. The heat maps demonstrated the effect of BAY 11-7082 in gene expression of NF-κB pathway and NF-κB responsive genes (red color for maximum expression and green for minimum). Group 1: Acidic bile-treated cells with BAY 11-7082; Control Group: Acidic bile-treated cells without BAY 11-7082. (Gene expression has been normalized to two housekeeping genes; hGAPDH, human <t>glyceraldehyde-3-phosphate</t> dehydrogenase and RPLP0, ribosomal protein lateral stalk subunit P0).
    Human Glyceraldehyde 3 Phosphate Dehydrogenase, supplied by Qiagen, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 92 stars, based on 1 article reviews
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    85
    Qiagen mouse glyceraldehyde 3 phosphate dehydrogenase gapdh cdna measurements
    mRNA expression levels of the target gene in various organs . mRNA levels were evaluated using real time RT-PCR. Plasmid DNA (100 μg) complexed with arginine peptide at an N/P ratio of 3:1 was intraperitoneally administered to mice. Mice were sacrificed at the indicated time points, and total RNA was extracted from the organs. After preparation of <t>cDNA,</t> PCR amplification of luciferase and <t>GAPDH</t> genes was performed using the specific primers described in the Materials and Methods section. Results are expressed as means ± S.D. for at least 3 different experiments.
    Mouse Glyceraldehyde 3 Phosphate Dehydrogenase Gapdh Cdna Measurements, supplied by Qiagen, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Hp reduces Hb-induced cellular activation, inflammatory response and oxidative stress in the choroid plexus. Rabbit pups with confirmed IVH, injected intraventricularly with Hp (grey bars; n = 6) or sham (white bars; n = 6) 6 hours after confirmation of the bleeding, were euthanized at 24 hours of age and the brains were removed from the skulls and the choroid plexus was carefully removed from the lateral ventricles, snap frozen, and the mRNA expression of IL1R1 (A) , FAS (B) , NF-Κβ (C) , MCP-1 (D) , IL-8 (E) , IL-1β (F) , TNFα (G) , IL-6 (H) and HO-1 (I) were subsequently analyzed with real-time PCR, as described in Materials and Methods. mRNA expression for respective gene was normalized against those of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against samples from control pups. Results are presented as box plots displaying medians and 25 th and 75 th percentiles. Differences between IVH + Hp versus IVH + Sham at 24 hours were analyzed using the Mann-Whitney U -test. * P

    Journal: Journal of Neuroinflammation

    Article Title: Extracellular hemoglobin - mediator of inflammation and cell death in the choroid plexus following preterm intraventricular hemorrhage

    doi: 10.1186/s12974-014-0200-9

    Figure Lengend Snippet: Hp reduces Hb-induced cellular activation, inflammatory response and oxidative stress in the choroid plexus. Rabbit pups with confirmed IVH, injected intraventricularly with Hp (grey bars; n = 6) or sham (white bars; n = 6) 6 hours after confirmation of the bleeding, were euthanized at 24 hours of age and the brains were removed from the skulls and the choroid plexus was carefully removed from the lateral ventricles, snap frozen, and the mRNA expression of IL1R1 (A) , FAS (B) , NF-Κβ (C) , MCP-1 (D) , IL-8 (E) , IL-1β (F) , TNFα (G) , IL-6 (H) and HO-1 (I) were subsequently analyzed with real-time PCR, as described in Materials and Methods. mRNA expression for respective gene was normalized against those of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against samples from control pups. Results are presented as box plots displaying medians and 25 th and 75 th percentiles. Differences between IVH + Hp versus IVH + Sham at 24 hours were analyzed using the Mann-Whitney U -test. * P

    Article Snippet: Data were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH, custom made by QIAGEN; Germantown, MD, USA).

    Techniques: Activation Assay, Injection, Expressing, Real-time Polymerase Chain Reaction, MANN-WHITNEY

    Hp reduces Hb-induced cellular activation, inflammatory response and oxidative stress in choroid plexus epithelial cell cultures. mRNA expression of NF-Κβ (A) , MCP-1 (B) , IL-8 (C) , IL-1β (D) , IL-6 (E) and HO-1 (F) in HCPEpiC cells, exposed to 10% (of cell culture medium volume) cerebrospinal fluid (CSF) from preterm infants with IVH or 10 μM metHb with or without the addition of 1.0 mg/ml Hp, for 4 and 24 hours was determined using real-time PCR as described in Materials and Methods. The mRNA expression of all respective genes was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. Differences between the respective exposures versus control for respective time point were analyzed using ANOVA post hoc Bonferroni. * P

    Journal: Journal of Neuroinflammation

    Article Title: Extracellular hemoglobin - mediator of inflammation and cell death in the choroid plexus following preterm intraventricular hemorrhage

    doi: 10.1186/s12974-014-0200-9

    Figure Lengend Snippet: Hp reduces Hb-induced cellular activation, inflammatory response and oxidative stress in choroid plexus epithelial cell cultures. mRNA expression of NF-Κβ (A) , MCP-1 (B) , IL-8 (C) , IL-1β (D) , IL-6 (E) and HO-1 (F) in HCPEpiC cells, exposed to 10% (of cell culture medium volume) cerebrospinal fluid (CSF) from preterm infants with IVH or 10 μM metHb with or without the addition of 1.0 mg/ml Hp, for 4 and 24 hours was determined using real-time PCR as described in Materials and Methods. The mRNA expression of all respective genes was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. Differences between the respective exposures versus control for respective time point were analyzed using ANOVA post hoc Bonferroni. * P

    Article Snippet: Data were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH, custom made by QIAGEN; Germantown, MD, USA).

    Techniques: Activation Assay, Expressing, Cell Culture, Real-time Polymerase Chain Reaction

    CSF-induced cellular activation, inflammatory response and oxidative stress in choroid plexus epithelial cell cultures. mRNA expression of NF-Κβ (A) , MCP-1 (B) , IL-8 (C) , IL-1β (D) , IL-6 (E) and HO-1 (F) in HCPEpiC cells, exposed to culture medium containing 1 to 30% (of cell culture medium volume) CSF from preterm infants with intraventricular hemorrhage (IVH) for 4 and 24 hours was determined using real-time PCR as described in Materials and Methods. The mRNA expression of all respective genes was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. Differences between the respective exposures versus control for respective time point were analyzed using ANOVA post hoc Bonferroni. * P

    Journal: Journal of Neuroinflammation

    Article Title: Extracellular hemoglobin - mediator of inflammation and cell death in the choroid plexus following preterm intraventricular hemorrhage

    doi: 10.1186/s12974-014-0200-9

    Figure Lengend Snippet: CSF-induced cellular activation, inflammatory response and oxidative stress in choroid plexus epithelial cell cultures. mRNA expression of NF-Κβ (A) , MCP-1 (B) , IL-8 (C) , IL-1β (D) , IL-6 (E) and HO-1 (F) in HCPEpiC cells, exposed to culture medium containing 1 to 30% (of cell culture medium volume) CSF from preterm infants with intraventricular hemorrhage (IVH) for 4 and 24 hours was determined using real-time PCR as described in Materials and Methods. The mRNA expression of all respective genes was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. Differences between the respective exposures versus control for respective time point were analyzed using ANOVA post hoc Bonferroni. * P

    Article Snippet: Data were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH, custom made by QIAGEN; Germantown, MD, USA).

    Techniques: Activation Assay, Expressing, Cell Culture, Real-time Polymerase Chain Reaction

    MetHb- and heme-induced cellular activation, inflammatory response and oxidative stress in choroid plexus epithelial cell cultures. mRNA expression of NF-Κβ (A) , MCP-1 (B) , IL-8 (C) , IL-1β (D) , IL-6 (E) and HO-1 (F) in HCPEpiC cells, exposed to 10 μM metHb or heme for 4 and 24 hours was determined using real-time PCR as described in Materials and Methods. The mRNA expression of all respective genes was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. Differences between the respective exposures versus control for respective time point were analyzed using ANOVA post hoc Bonferroni. * P

    Journal: Journal of Neuroinflammation

    Article Title: Extracellular hemoglobin - mediator of inflammation and cell death in the choroid plexus following preterm intraventricular hemorrhage

    doi: 10.1186/s12974-014-0200-9

    Figure Lengend Snippet: MetHb- and heme-induced cellular activation, inflammatory response and oxidative stress in choroid plexus epithelial cell cultures. mRNA expression of NF-Κβ (A) , MCP-1 (B) , IL-8 (C) , IL-1β (D) , IL-6 (E) and HO-1 (F) in HCPEpiC cells, exposed to 10 μM metHb or heme for 4 and 24 hours was determined using real-time PCR as described in Materials and Methods. The mRNA expression of all respective genes was normalized against glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and is given as fold change. The fold-change values were calculated by normalizing against control samples from untreated cells. Results are from triplicate experiments and presented as mean ± SEM. Differences between the respective exposures versus control for respective time point were analyzed using ANOVA post hoc Bonferroni. * P

    Article Snippet: Data were normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH, custom made by QIAGEN; Germantown, MD, USA).

    Techniques: Activation Assay, Expressing, Real-time Polymerase Chain Reaction

    Real-time PCR gene expression comparison between BM testes and germ cell-derived colonies. Relative gene expressions were compared between GDCs and 4-month-old BM testes.  GAPDH , glyceraldehyde 3-phosphate dehydrogenase;  PGP9.5 , protein gene product 9.5 (alternate designation: ubiquitin carboxy-terminal hydrolase L1);  GFRα-1 , glial cell line derived neurotrophic factor family receptor alpha 1;  PLZF , promyelocytic leukaemia zinc finger;  Oct4 , octamer-binding protein 4;  Nanog , homeobox transcription factor Nanog;  GATA4 , GATA-binding protein 4. (* P

    Journal: Scientific Reports

    Article Title: Vitrified canine testicular cells allow the formation of spermatogonial stem cells and seminiferous tubules following their xenotransplantation into nude mice

    doi: 10.1038/srep21919

    Figure Lengend Snippet: Real-time PCR gene expression comparison between BM testes and germ cell-derived colonies. Relative gene expressions were compared between GDCs and 4-month-old BM testes. GAPDH , glyceraldehyde 3-phosphate dehydrogenase; PGP9.5 , protein gene product 9.5 (alternate designation: ubiquitin carboxy-terminal hydrolase L1); GFRα-1 , glial cell line derived neurotrophic factor family receptor alpha 1; PLZF , promyelocytic leukaemia zinc finger; Oct4 , octamer-binding protein 4; Nanog , homeobox transcription factor Nanog; GATA4 , GATA-binding protein 4. (* P

    Article Snippet: Real-time PCR was performed with glyceraldehyde 3-phosphate dehydrogenase (GAPDH ), GFRα-1 , PLZF , Oct4 , Nanog , and GATA-binding protein 4 (GATA4 ) primers, using Rotor-Gene Q (Qiagen, Venlo, the Netherlands).

    Techniques: Real-time Polymerase Chain Reaction, Expressing, Derivative Assay, Binding Assay

    BAY 11-7082 reduced the acidic bile-induced gene expression profiling of NF-κB signaling pathway in treated normal human hypopharyngeal cells Heat maps were obtained by RT 2 -Profiler PCR array analysis for NF-κB signaling. The genes were clustered based on their biological role. The heat maps demonstrated the effect of BAY 11-7082 in gene expression of NF-κB pathway and NF-κB responsive genes (red color for maximum expression and green for minimum). Group 1: Acidic bile-treated cells with BAY 11-7082; Control Group: Acidic bile-treated cells without BAY 11-7082. (Gene expression has been normalized to two housekeeping genes; hGAPDH, human glyceraldehyde-3-phosphate dehydrogenase and RPLP0, ribosomal protein lateral stalk subunit P0).

    Journal: Oncotarget

    Article Title: Inhibition of NF-κB prevents the acidic bile-induced oncogenic mRNA phenotype, in human hypopharyngeal cells

    doi: 10.18632/oncotarget.23143

    Figure Lengend Snippet: BAY 11-7082 reduced the acidic bile-induced gene expression profiling of NF-κB signaling pathway in treated normal human hypopharyngeal cells Heat maps were obtained by RT 2 -Profiler PCR array analysis for NF-κB signaling. The genes were clustered based on their biological role. The heat maps demonstrated the effect of BAY 11-7082 in gene expression of NF-κB pathway and NF-κB responsive genes (red color for maximum expression and green for minimum). Group 1: Acidic bile-treated cells with BAY 11-7082; Control Group: Acidic bile-treated cells without BAY 11-7082. (Gene expression has been normalized to two housekeeping genes; hGAPDH, human glyceraldehyde-3-phosphate dehydrogenase and RPLP0, ribosomal protein lateral stalk subunit P0).

    Article Snippet: We performed reverse transcription (iScript cDNA synthesis kit; Bio-Rad) and real time qPCR analysis (Bio-Rad real time thermal cycler CFX96TM; Bio-Rad) using specific primers for target genes and reference housekeeping gene, human glyceraldehyde 3-phosphate dehydrogenase (h GAPDH) ( ; see online), (QuantiTect Primers Assays; Qiagen), and iQ™ SYBR Green Supermix (Bio-Rad).

    Techniques: Expressing, Polymerase Chain Reaction

    mRNA expression levels of the target gene in various organs . mRNA levels were evaluated using real time RT-PCR. Plasmid DNA (100 μg) complexed with arginine peptide at an N/P ratio of 3:1 was intraperitoneally administered to mice. Mice were sacrificed at the indicated time points, and total RNA was extracted from the organs. After preparation of cDNA, PCR amplification of luciferase and GAPDH genes was performed using the specific primers described in the Materials and Methods section. Results are expressed as means ± S.D. for at least 3 different experiments.

    Journal: Genetic Vaccines and Therapy

    Article Title: Biodistribution and blood clearance of plasmid DNA administered in arginine peptide complexes

    doi: 10.1186/1479-0556-9-13

    Figure Lengend Snippet: mRNA expression levels of the target gene in various organs . mRNA levels were evaluated using real time RT-PCR. Plasmid DNA (100 μg) complexed with arginine peptide at an N/P ratio of 3:1 was intraperitoneally administered to mice. Mice were sacrificed at the indicated time points, and total RNA was extracted from the organs. After preparation of cDNA, PCR amplification of luciferase and GAPDH genes was performed using the specific primers described in the Materials and Methods section. Results are expressed as means ± S.D. for at least 3 different experiments.

    Article Snippet: The thermal cycler protocol was set as follows: pre-incubation at 95°C for 10 s, amplification at 40 cycles at 95°C for 5 s, and 60°C for 40 s. For the mouse glyceraldehyde-3-phosphate dehydrogenase (GAPDH) cDNA measurements, each sample was prepared following the manufacturer's instructions with a GAPDH primer set (Qiagen).

    Techniques: Expressing, Quantitative RT-PCR, Plasmid Preparation, Mouse Assay, Polymerase Chain Reaction, Amplification, Luciferase

    Effects of DNA dose on plasmid DNA expression after delivery in arginine/DNA complexes . Various amounts of plasmid DNA complexed with arginine peptide at an N/P ratio of 3:1 were intraperitoneally administered to mice, and mRNA levels were evaluated using real time RT-PCR. Total RNA was extracted from the organs. After preparation of cDNA, PCR amplification of luciferase and GAPDH genes was performed using the specific primers described in the Materials and Methods section. Results are expressed as means ± S.D. for at least 3 different experiments.

    Journal: Genetic Vaccines and Therapy

    Article Title: Biodistribution and blood clearance of plasmid DNA administered in arginine peptide complexes

    doi: 10.1186/1479-0556-9-13

    Figure Lengend Snippet: Effects of DNA dose on plasmid DNA expression after delivery in arginine/DNA complexes . Various amounts of plasmid DNA complexed with arginine peptide at an N/P ratio of 3:1 were intraperitoneally administered to mice, and mRNA levels were evaluated using real time RT-PCR. Total RNA was extracted from the organs. After preparation of cDNA, PCR amplification of luciferase and GAPDH genes was performed using the specific primers described in the Materials and Methods section. Results are expressed as means ± S.D. for at least 3 different experiments.

    Article Snippet: The thermal cycler protocol was set as follows: pre-incubation at 95°C for 10 s, amplification at 40 cycles at 95°C for 5 s, and 60°C for 40 s. For the mouse glyceraldehyde-3-phosphate dehydrogenase (GAPDH) cDNA measurements, each sample was prepared following the manufacturer's instructions with a GAPDH primer set (Qiagen).

    Techniques: Plasmid Preparation, Expressing, Mouse Assay, Quantitative RT-PCR, Polymerase Chain Reaction, Amplification, Luciferase

    Duration of plasmid DNA expression . Plasmid DNA (100 μg) complexed with arginine peptide at an N/P ratio of 3:1 was intraperitoneally administered to mice, and total RNA was extracted from the organs at the indicated time points. The RNA extracts were transformed to cDNA using RT-PCR to serve as templates for nested PCR analysis. PCR amplification of luciferase and GAPDH genes was performed using the specific primers described in the Materials and Methods section. The nested PCR products were separated on a 1.2% agarose gel.

    Journal: Genetic Vaccines and Therapy

    Article Title: Biodistribution and blood clearance of plasmid DNA administered in arginine peptide complexes

    doi: 10.1186/1479-0556-9-13

    Figure Lengend Snippet: Duration of plasmid DNA expression . Plasmid DNA (100 μg) complexed with arginine peptide at an N/P ratio of 3:1 was intraperitoneally administered to mice, and total RNA was extracted from the organs at the indicated time points. The RNA extracts were transformed to cDNA using RT-PCR to serve as templates for nested PCR analysis. PCR amplification of luciferase and GAPDH genes was performed using the specific primers described in the Materials and Methods section. The nested PCR products were separated on a 1.2% agarose gel.

    Article Snippet: The thermal cycler protocol was set as follows: pre-incubation at 95°C for 10 s, amplification at 40 cycles at 95°C for 5 s, and 60°C for 40 s. For the mouse glyceraldehyde-3-phosphate dehydrogenase (GAPDH) cDNA measurements, each sample was prepared following the manufacturer's instructions with a GAPDH primer set (Qiagen).

    Techniques: Plasmid Preparation, Expressing, Mouse Assay, Transformation Assay, Reverse Transcription Polymerase Chain Reaction, Nested PCR, Polymerase Chain Reaction, Amplification, Luciferase, Agarose Gel Electrophoresis