nerve growth factor ngf  (Alomone Labs)


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    Structured Review

    Alomone Labs nerve growth factor ngf
    <t>BDNF-induced</t> NR2C upregulation by TrkB activation. Cells were cultured in low KCl and treated with or without BDNF (100 ng/ml in A and 50 ng/ml in B-D and F ) for 96 h. A , Levels of indicated mRNAs were quantitated by RNA blotting ( n = 4). B , Cell lysates (40 μg) were immunoblotted with anti-panNR2 antibody or anti-NR1 antibody. Molecular sizes (kilodaltons) of protein makers are indicated on the left. C , P2 membrane fractions were isolated, solubilized, and immunoprecipitated (IP) with anti-NR1 antibody, followed by immnoblotting with anti-panNR2 antibody. D , Cell-surface proteins were biotinylated with Sulfo-NHS-SS-Biotin. Cell lysates were solubilized, precipitated with NeutrAvidin beads, and immunoblotted with anti-panNR2 antibody. E , Cells were cultured in low KCl and treated with BDNF, <t>NGF,</t> NT-3, or NT-4 (50 ng/ml each) for 96 h. Levels of NR2C mRNA were quantitated ( n = 4). F , Granule cells were prepared from TrkB - / - knock-out mice(KO) and their wild-type (WT) littermates. NR2C and TrkB mRNAs were analyzed by RNA blotting. Ethidium bromide-stained 18s rRNA is also indicated. * p
    Nerve Growth Factor Ngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 30 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "Neuronal Depolarization Controls Brain-Derived Neurotrophic Factor-Induced Upregulation of NR2C NMDA Receptor via Calcineurin Signaling"

    Article Title: Neuronal Depolarization Controls Brain-Derived Neurotrophic Factor-Induced Upregulation of NR2C NMDA Receptor via Calcineurin Signaling

    Journal: The Journal of Neuroscience

    doi: 10.1523/JNEUROSCI.2191-05.2005

    BDNF-induced NR2C upregulation by TrkB activation. Cells were cultured in low KCl and treated with or without BDNF (100 ng/ml in A and 50 ng/ml in B-D and F ) for 96 h. A , Levels of indicated mRNAs were quantitated by RNA blotting ( n = 4). B , Cell lysates (40 μg) were immunoblotted with anti-panNR2 antibody or anti-NR1 antibody. Molecular sizes (kilodaltons) of protein makers are indicated on the left. C , P2 membrane fractions were isolated, solubilized, and immunoprecipitated (IP) with anti-NR1 antibody, followed by immnoblotting with anti-panNR2 antibody. D , Cell-surface proteins were biotinylated with Sulfo-NHS-SS-Biotin. Cell lysates were solubilized, precipitated with NeutrAvidin beads, and immunoblotted with anti-panNR2 antibody. E , Cells were cultured in low KCl and treated with BDNF, NGF, NT-3, or NT-4 (50 ng/ml each) for 96 h. Levels of NR2C mRNA were quantitated ( n = 4). F , Granule cells were prepared from TrkB - / - knock-out mice(KO) and their wild-type (WT) littermates. NR2C and TrkB mRNAs were analyzed by RNA blotting. Ethidium bromide-stained 18s rRNA is also indicated. * p
    Figure Legend Snippet: BDNF-induced NR2C upregulation by TrkB activation. Cells were cultured in low KCl and treated with or without BDNF (100 ng/ml in A and 50 ng/ml in B-D and F ) for 96 h. A , Levels of indicated mRNAs were quantitated by RNA blotting ( n = 4). B , Cell lysates (40 μg) were immunoblotted with anti-panNR2 antibody or anti-NR1 antibody. Molecular sizes (kilodaltons) of protein makers are indicated on the left. C , P2 membrane fractions were isolated, solubilized, and immunoprecipitated (IP) with anti-NR1 antibody, followed by immnoblotting with anti-panNR2 antibody. D , Cell-surface proteins were biotinylated with Sulfo-NHS-SS-Biotin. Cell lysates were solubilized, precipitated with NeutrAvidin beads, and immunoblotted with anti-panNR2 antibody. E , Cells were cultured in low KCl and treated with BDNF, NGF, NT-3, or NT-4 (50 ng/ml each) for 96 h. Levels of NR2C mRNA were quantitated ( n = 4). F , Granule cells were prepared from TrkB - / - knock-out mice(KO) and their wild-type (WT) littermates. NR2C and TrkB mRNAs were analyzed by RNA blotting. Ethidium bromide-stained 18s rRNA is also indicated. * p

    Techniques Used: Activation Assay, Cell Culture, Isolation, Immunoprecipitation, Knock-Out, Mouse Assay, Staining

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    Alomone Labs recombinant rat ngf
    <t>TGF-β1</t> promoted the mRNA expression of <t>NGF</t> in SCDC2 cells through its type I receptor in a dose-dependent manner. After 24-h culture in growth medium, SCDC2 cells were starved for 24 h. The starved cells were then treated with (A) TGF-β1 at various concentrations for 24 h, or (B) pretreated with or without TGF-β type I receptor inhibitor SB-431542 (10 µ M) for 30 min and then with or without TGF-β1 (10 ng/ml) for 24 h. (C) Starved cells were treated with or without TGF-β1 (10 ng/ml) for the indicated times. The relative expression level of NGF was evaluated using reverse transcription-quantitative polymerase chain reaction. Data represent the mean ± standard deviation (n=6). * P
    Recombinant Rat Ngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs pc12 cells
    Irradiation with 137 Cs γ-rays depresses NGF-induced neurite extension in <t>PC12</t> cells. (a) Phase-contrast micrographs, (b) lengths of neurites and (c) numbers of neurites in PC12 cells after 5 days of NGF stimulation or non-stimulation, with or without 137 Csγ-ray irradiation. Lengths or numbers of neurites are expressed as the relative ratio to the non-irradiated group. Data are presented as the mean ± standard error of triplicate samples. * P
    Pc12 Cells, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Alomone Labs nerve growth factor ngf
    <t>BDNF-induced</t> NR2C upregulation by TrkB activation. Cells were cultured in low KCl and treated with or without BDNF (100 ng/ml in A and 50 ng/ml in B-D and F ) for 96 h. A , Levels of indicated mRNAs were quantitated by RNA blotting ( n = 4). B , Cell lysates (40 μg) were immunoblotted with anti-panNR2 antibody or anti-NR1 antibody. Molecular sizes (kilodaltons) of protein makers are indicated on the left. C , P2 membrane fractions were isolated, solubilized, and immunoprecipitated (IP) with anti-NR1 antibody, followed by immnoblotting with anti-panNR2 antibody. D , Cell-surface proteins were biotinylated with Sulfo-NHS-SS-Biotin. Cell lysates were solubilized, precipitated with NeutrAvidin beads, and immunoblotted with anti-panNR2 antibody. E , Cells were cultured in low KCl and treated with BDNF, <t>NGF,</t> NT-3, or NT-4 (50 ng/ml each) for 96 h. Levels of NR2C mRNA were quantitated ( n = 4). F , Granule cells were prepared from TrkB - / - knock-out mice(KO) and their wild-type (WT) littermates. NR2C and TrkB mRNAs were analyzed by RNA blotting. Ethidium bromide-stained 18s rRNA is also indicated. * p
    Nerve Growth Factor Ngf, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nerve growth factor ngf/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    nerve growth factor ngf - by Bioz Stars, 2022-08
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    Image Search Results


    TGF-β1 promoted the mRNA expression of NGF in SCDC2 cells through its type I receptor in a dose-dependent manner. After 24-h culture in growth medium, SCDC2 cells were starved for 24 h. The starved cells were then treated with (A) TGF-β1 at various concentrations for 24 h, or (B) pretreated with or without TGF-β type I receptor inhibitor SB-431542 (10 µ M) for 30 min and then with or without TGF-β1 (10 ng/ml) for 24 h. (C) Starved cells were treated with or without TGF-β1 (10 ng/ml) for the indicated times. The relative expression level of NGF was evaluated using reverse transcription-quantitative polymerase chain reaction. Data represent the mean ± standard deviation (n=6). * P

    Journal: International Journal of Molecular Medicine

    Article Title: IL-1β and TNF-α suppress TGF-β-promoted NGF expression in periodontal ligament-derived fibroblasts through inactivation of TGF-β-induced Smad2/3- and p38 MAPK-mediated signals

    doi: 10.3892/ijmm_2018.3714

    Figure Lengend Snippet: TGF-β1 promoted the mRNA expression of NGF in SCDC2 cells through its type I receptor in a dose-dependent manner. After 24-h culture in growth medium, SCDC2 cells were starved for 24 h. The starved cells were then treated with (A) TGF-β1 at various concentrations for 24 h, or (B) pretreated with or without TGF-β type I receptor inhibitor SB-431542 (10 µ M) for 30 min and then with or without TGF-β1 (10 ng/ml) for 24 h. (C) Starved cells were treated with or without TGF-β1 (10 ng/ml) for the indicated times. The relative expression level of NGF was evaluated using reverse transcription-quantitative polymerase chain reaction. Data represent the mean ± standard deviation (n=6). * P

    Article Snippet: Recombinant human TGF-β1 was obtained from PeproTech, Inc. (Rocky Hill, NJ, USA), recombinant rat NGF was obtained from Alomone Labs (Jerusalem, Israel), and recombinant rat IL-1β and TNF-α were purchased from Miltenyi Biotec GmbH (Bergisch Gladbach, Germany).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Standard Deviation

    IL-1β and TNF-α suppressed the TGF-β1-induced mRNA expression of NGF in SCDC2 cells by abrogating Smad2/3 and p38 MAPK activities. The effects of IL-1β and TNF-α on TGF-β1-induced mRNA expression of NGF in SCDC2 cells were evaluated using RT-qPCR. The cells were treated with or without (A) IL-1β alone or (B) TNF-α alone at indicated concentrations, (C) TGF-β1 (10 ng/ml) and/or IL-1β (10 ng/ml), and (D) TGF-β1 (10 ng/ml) and/or TNF-α (10 ng/ml). Data represent the mean ± standard deviation (n=6). * P

    Journal: International Journal of Molecular Medicine

    Article Title: IL-1β and TNF-α suppress TGF-β-promoted NGF expression in periodontal ligament-derived fibroblasts through inactivation of TGF-β-induced Smad2/3- and p38 MAPK-mediated signals

    doi: 10.3892/ijmm_2018.3714

    Figure Lengend Snippet: IL-1β and TNF-α suppressed the TGF-β1-induced mRNA expression of NGF in SCDC2 cells by abrogating Smad2/3 and p38 MAPK activities. The effects of IL-1β and TNF-α on TGF-β1-induced mRNA expression of NGF in SCDC2 cells were evaluated using RT-qPCR. The cells were treated with or without (A) IL-1β alone or (B) TNF-α alone at indicated concentrations, (C) TGF-β1 (10 ng/ml) and/or IL-1β (10 ng/ml), and (D) TGF-β1 (10 ng/ml) and/or TNF-α (10 ng/ml). Data represent the mean ± standard deviation (n=6). * P

    Article Snippet: Recombinant human TGF-β1 was obtained from PeproTech, Inc. (Rocky Hill, NJ, USA), recombinant rat NGF was obtained from Alomone Labs (Jerusalem, Israel), and recombinant rat IL-1β and TNF-α were purchased from Miltenyi Biotec GmbH (Bergisch Gladbach, Germany).

    Techniques: Expressing, Quantitative RT-PCR, Standard Deviation

    TGF-β1 promoted the mRNA expression of NGF in SCDC2 cells in Smad2/3-dependent and p38 MAPK-dependent manners. Effects of (A) SIS3 (10 µ M), and (B) SB203580 (10 µ M) on expression of NGF mRNA were evaluated as described in Materials and methods. Data represent the mean ± standard deviation (n=6). * P

    Journal: International Journal of Molecular Medicine

    Article Title: IL-1β and TNF-α suppress TGF-β-promoted NGF expression in periodontal ligament-derived fibroblasts through inactivation of TGF-β-induced Smad2/3- and p38 MAPK-mediated signals

    doi: 10.3892/ijmm_2018.3714

    Figure Lengend Snippet: TGF-β1 promoted the mRNA expression of NGF in SCDC2 cells in Smad2/3-dependent and p38 MAPK-dependent manners. Effects of (A) SIS3 (10 µ M), and (B) SB203580 (10 µ M) on expression of NGF mRNA were evaluated as described in Materials and methods. Data represent the mean ± standard deviation (n=6). * P

    Article Snippet: Recombinant human TGF-β1 was obtained from PeproTech, Inc. (Rocky Hill, NJ, USA), recombinant rat NGF was obtained from Alomone Labs (Jerusalem, Israel), and recombinant rat IL-1β and TNF-α were purchased from Miltenyi Biotec GmbH (Bergisch Gladbach, Germany).

    Techniques: Expressing, Standard Deviation

    Irradiation with 137 Cs γ-rays depresses NGF-induced neurite extension in PC12 cells. (a) Phase-contrast micrographs, (b) lengths of neurites and (c) numbers of neurites in PC12 cells after 5 days of NGF stimulation or non-stimulation, with or without 137 Csγ-ray irradiation. Lengths or numbers of neurites are expressed as the relative ratio to the non-irradiated group. Data are presented as the mean ± standard error of triplicate samples. * P

    Journal: Journal of Radiation Research

    Article Title: Chronic irradiation with low-dose-rate 137Cs-γ rays inhibits NGF-induced neurite extension of PC12 cells via Ca2+/calmodulin-dependent kinase II activation

    doi: 10.1093/jrr/rrx032

    Figure Lengend Snippet: Irradiation with 137 Cs γ-rays depresses NGF-induced neurite extension in PC12 cells. (a) Phase-contrast micrographs, (b) lengths of neurites and (c) numbers of neurites in PC12 cells after 5 days of NGF stimulation or non-stimulation, with or without 137 Csγ-ray irradiation. Lengths or numbers of neurites are expressed as the relative ratio to the non-irradiated group. Data are presented as the mean ± standard error of triplicate samples. * P

    Article Snippet: Reagents We utilized the following reagents to investigate the effects of chronic, low-dose-rate 137 Cs-γ radiation on NGF-induced neurite extension in PC12 cells: NGF 2.5S (N-100, Alomone Labs, Jerusalem, Israel); anti-ERK1/ERK2 mouse-monoclonal antibody (MAB1576) and anti-phosphorylated ERK1/ERK2 rabbit-polyclonal antibody (MAB1018, both R & D systems, Minneapolis, MN, USA); anti-Trk A rabbit polyclonal antibody (sc-118) and anti-phosphorylated Trk A mouse-monoclonal antibody (sc-8058, both Santa Cruz Biotechnology, Dallas, TX, USA)—Trk A is a high-affinity nerve growth factor receptor; anti-tyrosine hydroxylase rabbit-monoclonal antibody (ab137869); anti-Akt rabbit polyclonal antibody (ab8805) and anti-phosphorylated Akt rabbit monoclonal antibody (ab81283); anti-Ca2+/calmodulin-dependent kinase II (CaMKII) rabbit polyclonal antibody (ab131468) and anti-phosphorylated CaMKII rabbit polyclonal antibody (ab5683, all Abcam plc, Cambridge, UK); KN-62 (I2142, Sigma-Aldrich, St Louis, MO, USA).

    Techniques: Irradiation

    Western blot analysis of NGF stimulation–related proteins in PC12 cells. Immunoblot showing varying levels of (a) phosphorylated NGF receptor (P-NGFR, upper) and NGF receptor (NGFR, lower), (b) phosphorylated ERK (P-ERK, upper) and ERK (lower), (c) tyrosine hydroxylase (TH) observed in PC12 cells after 5 days of NGF stimulation or non-stimulation, with or without 137 Csγ-ray irradiation. Similar results were obtained in three separate experiments. NGF = nerve growth factor, ERK = extracellular signal–regulated kinase.

    Journal: Journal of Radiation Research

    Article Title: Chronic irradiation with low-dose-rate 137Cs-γ rays inhibits NGF-induced neurite extension of PC12 cells via Ca2+/calmodulin-dependent kinase II activation

    doi: 10.1093/jrr/rrx032

    Figure Lengend Snippet: Western blot analysis of NGF stimulation–related proteins in PC12 cells. Immunoblot showing varying levels of (a) phosphorylated NGF receptor (P-NGFR, upper) and NGF receptor (NGFR, lower), (b) phosphorylated ERK (P-ERK, upper) and ERK (lower), (c) tyrosine hydroxylase (TH) observed in PC12 cells after 5 days of NGF stimulation or non-stimulation, with or without 137 Csγ-ray irradiation. Similar results were obtained in three separate experiments. NGF = nerve growth factor, ERK = extracellular signal–regulated kinase.

    Article Snippet: Reagents We utilized the following reagents to investigate the effects of chronic, low-dose-rate 137 Cs-γ radiation on NGF-induced neurite extension in PC12 cells: NGF 2.5S (N-100, Alomone Labs, Jerusalem, Israel); anti-ERK1/ERK2 mouse-monoclonal antibody (MAB1576) and anti-phosphorylated ERK1/ERK2 rabbit-polyclonal antibody (MAB1018, both R & D systems, Minneapolis, MN, USA); anti-Trk A rabbit polyclonal antibody (sc-118) and anti-phosphorylated Trk A mouse-monoclonal antibody (sc-8058, both Santa Cruz Biotechnology, Dallas, TX, USA)—Trk A is a high-affinity nerve growth factor receptor; anti-tyrosine hydroxylase rabbit-monoclonal antibody (ab137869); anti-Akt rabbit polyclonal antibody (ab8805) and anti-phosphorylated Akt rabbit monoclonal antibody (ab81283); anti-Ca2+/calmodulin-dependent kinase II (CaMKII) rabbit polyclonal antibody (ab131468) and anti-phosphorylated CaMKII rabbit polyclonal antibody (ab5683, all Abcam plc, Cambridge, UK); KN-62 (I2142, Sigma-Aldrich, St Louis, MO, USA).

    Techniques: Western Blot, Irradiation

    Inhibition of CaMKII activity results in 137 Csγ-ray irradiation-induced depression of NGF-induced neurite extension in PC12 cells. (a) Immunoblot showing varying levels of phosphorylated CaMKII (P-CaMKII, upper) and CaMKII (lower) observed in PC12 cells after 5 days of NGF stimulation or non-stimulation, with or without 137 Csγ-ray irradiation in the presence or absence of KN-62. Similar results were obtained in three separate experiments. (b) Lengths and (c) numbers of neurites in PC12 cells after 5 days of NGF stimulation or non-stimulation, with or without 137 Csγ-ray irradiation in the presence or absence of KN-62. Lengths or numbers of neurites are expressed as the relative ratio to the non-irradiated group. Data are presented as the mean ± standard error of triplicate samples. * P

    Journal: Journal of Radiation Research

    Article Title: Chronic irradiation with low-dose-rate 137Cs-γ rays inhibits NGF-induced neurite extension of PC12 cells via Ca2+/calmodulin-dependent kinase II activation

    doi: 10.1093/jrr/rrx032

    Figure Lengend Snippet: Inhibition of CaMKII activity results in 137 Csγ-ray irradiation-induced depression of NGF-induced neurite extension in PC12 cells. (a) Immunoblot showing varying levels of phosphorylated CaMKII (P-CaMKII, upper) and CaMKII (lower) observed in PC12 cells after 5 days of NGF stimulation or non-stimulation, with or without 137 Csγ-ray irradiation in the presence or absence of KN-62. Similar results were obtained in three separate experiments. (b) Lengths and (c) numbers of neurites in PC12 cells after 5 days of NGF stimulation or non-stimulation, with or without 137 Csγ-ray irradiation in the presence or absence of KN-62. Lengths or numbers of neurites are expressed as the relative ratio to the non-irradiated group. Data are presented as the mean ± standard error of triplicate samples. * P

    Article Snippet: Reagents We utilized the following reagents to investigate the effects of chronic, low-dose-rate 137 Cs-γ radiation on NGF-induced neurite extension in PC12 cells: NGF 2.5S (N-100, Alomone Labs, Jerusalem, Israel); anti-ERK1/ERK2 mouse-monoclonal antibody (MAB1576) and anti-phosphorylated ERK1/ERK2 rabbit-polyclonal antibody (MAB1018, both R & D systems, Minneapolis, MN, USA); anti-Trk A rabbit polyclonal antibody (sc-118) and anti-phosphorylated Trk A mouse-monoclonal antibody (sc-8058, both Santa Cruz Biotechnology, Dallas, TX, USA)—Trk A is a high-affinity nerve growth factor receptor; anti-tyrosine hydroxylase rabbit-monoclonal antibody (ab137869); anti-Akt rabbit polyclonal antibody (ab8805) and anti-phosphorylated Akt rabbit monoclonal antibody (ab81283); anti-Ca2+/calmodulin-dependent kinase II (CaMKII) rabbit polyclonal antibody (ab131468) and anti-phosphorylated CaMKII rabbit polyclonal antibody (ab5683, all Abcam plc, Cambridge, UK); KN-62 (I2142, Sigma-Aldrich, St Louis, MO, USA).

    Techniques: Inhibition, Activity Assay, Irradiation

    Irradiation with 137 Cs γ-rays attenuates NGF-induced Rac1 activation without increasing phosphorylation of Akt in PC12 cells. (a) The activity of Rac1 in PC12 cells after 5 days of NGF stimulation or non-stimulation, with or without 137 Csγ-ray irradiation. Rac1 activity is expressed as the relative ratio to the NGF non-stimulated group without irradiation. Data are presented as the mean ± standard error of triplicate samples. ** P

    Journal: Journal of Radiation Research

    Article Title: Chronic irradiation with low-dose-rate 137Cs-γ rays inhibits NGF-induced neurite extension of PC12 cells via Ca2+/calmodulin-dependent kinase II activation

    doi: 10.1093/jrr/rrx032

    Figure Lengend Snippet: Irradiation with 137 Cs γ-rays attenuates NGF-induced Rac1 activation without increasing phosphorylation of Akt in PC12 cells. (a) The activity of Rac1 in PC12 cells after 5 days of NGF stimulation or non-stimulation, with or without 137 Csγ-ray irradiation. Rac1 activity is expressed as the relative ratio to the NGF non-stimulated group without irradiation. Data are presented as the mean ± standard error of triplicate samples. ** P

    Article Snippet: Reagents We utilized the following reagents to investigate the effects of chronic, low-dose-rate 137 Cs-γ radiation on NGF-induced neurite extension in PC12 cells: NGF 2.5S (N-100, Alomone Labs, Jerusalem, Israel); anti-ERK1/ERK2 mouse-monoclonal antibody (MAB1576) and anti-phosphorylated ERK1/ERK2 rabbit-polyclonal antibody (MAB1018, both R & D systems, Minneapolis, MN, USA); anti-Trk A rabbit polyclonal antibody (sc-118) and anti-phosphorylated Trk A mouse-monoclonal antibody (sc-8058, both Santa Cruz Biotechnology, Dallas, TX, USA)—Trk A is a high-affinity nerve growth factor receptor; anti-tyrosine hydroxylase rabbit-monoclonal antibody (ab137869); anti-Akt rabbit polyclonal antibody (ab8805) and anti-phosphorylated Akt rabbit monoclonal antibody (ab81283); anti-Ca2+/calmodulin-dependent kinase II (CaMKII) rabbit polyclonal antibody (ab131468) and anti-phosphorylated CaMKII rabbit polyclonal antibody (ab5683, all Abcam plc, Cambridge, UK); KN-62 (I2142, Sigma-Aldrich, St Louis, MO, USA).

    Techniques: Irradiation, Activation Assay, Activity Assay

    SgG2 interacts with NGF. (A) Sensorgram showing the interaction of SgG2 with increasing concentrations of NGF. The concentrations of NGF are indicated at the right side of the sensorgram. (B) Sensorgram showing the interaction of SgG2 with NGF and the lack of interaction with TNF-α, IFN-α and IL-1. All analytes were injected at a 100 nM concentration. (C) Sensorgram depicting the interaction between increasing concentrations of SgG2 and NGF coupled on a sensor chip. As a negative control we used the same concentrations of purified HSV-2 gD. (D) Saturation curve for the binding of NGF to SgG2. The derived KD is shown (E) Sensorgram showing the interaction of coupled SgG2 with CXCL12β and NGF injected alone or in combination. In all cases the arrow indicates the end of injection. Abbreviations: Diff. Resp., Differential response; M, molar; R.U., response units; s, seconds.

    Journal: PLoS Pathogens

    Article Title: Secreted Herpes Simplex Virus-2 Glycoprotein G Modifies NGF-TrkA Signaling to Attract Free Nerve Endings to the Site of Infection

    doi: 10.1371/journal.ppat.1004571

    Figure Lengend Snippet: SgG2 interacts with NGF. (A) Sensorgram showing the interaction of SgG2 with increasing concentrations of NGF. The concentrations of NGF are indicated at the right side of the sensorgram. (B) Sensorgram showing the interaction of SgG2 with NGF and the lack of interaction with TNF-α, IFN-α and IL-1. All analytes were injected at a 100 nM concentration. (C) Sensorgram depicting the interaction between increasing concentrations of SgG2 and NGF coupled on a sensor chip. As a negative control we used the same concentrations of purified HSV-2 gD. (D) Saturation curve for the binding of NGF to SgG2. The derived KD is shown (E) Sensorgram showing the interaction of coupled SgG2 with CXCL12β and NGF injected alone or in combination. In all cases the arrow indicates the end of injection. Abbreviations: Diff. Resp., Differential response; M, molar; R.U., response units; s, seconds.

    Article Snippet: In brief, 340 μL of rat tail collagen I (BD Biosciences, San Jose, CA) were mixed with 40 μL of 10x MEM, 10 μL of HEPES or the vCKBPs at a final concentration of 50 nM, and mouse NGF 2.5S (N-100, Alomone labs, Jerusalem, Israel) at a final concentration of 0.25 nM.

    Techniques: Injection, Concentration Assay, Chromatin Immunoprecipitation, Negative Control, Purification, Binding Assay, Derivative Assay

    SgG2 reduces NGF induced TrkA internalization and retrograde transport. SCG dissociated neurons were grown during 5 DIV and deprived of NGF for 16 h. (A) Neurons were stimulated with NGF alone or together with the indicated viral proteins during 0, 15 and 120 min. TrkA internalization was analyzed by immunofluorescence without permeabilization and normalized for F-actin using phalloidin staining. Solid arrowhead points to a non-internalized cluster of TrkA at the plasma membrane. Images show a projection of at least 3 image planes. All images correspond to the same experiment, representative of two independent experiments ( n = 12 for each condition). Scale bar represents 5 μm. The graph shows the quantification of the TrkA/phalloidin staining at the plasma membrane over time in samples treated with NGF plus the indicated protein or buffer. (B) Mouse SCG dissociated neurons were grown during 7 DIV using microfluidic devices. Neurons were deprived of NGF for 16 h and NGF and viral proteins were added to the distal axon compartment during 120 min. TrkA phosphorylation was analyzed by immunofluorescence after permeabilization with Triton X-100 and normalized using phalloidin. The images and the graph correspond to the same experiment and are representative of three independent experiments. Scale bar 10 μm. n = 10 fields for each condition. A two-tailed unpaired T-test was used to calculate significance in (A) and (B) . *** P

    Journal: PLoS Pathogens

    Article Title: Secreted Herpes Simplex Virus-2 Glycoprotein G Modifies NGF-TrkA Signaling to Attract Free Nerve Endings to the Site of Infection

    doi: 10.1371/journal.ppat.1004571

    Figure Lengend Snippet: SgG2 reduces NGF induced TrkA internalization and retrograde transport. SCG dissociated neurons were grown during 5 DIV and deprived of NGF for 16 h. (A) Neurons were stimulated with NGF alone or together with the indicated viral proteins during 0, 15 and 120 min. TrkA internalization was analyzed by immunofluorescence without permeabilization and normalized for F-actin using phalloidin staining. Solid arrowhead points to a non-internalized cluster of TrkA at the plasma membrane. Images show a projection of at least 3 image planes. All images correspond to the same experiment, representative of two independent experiments ( n = 12 for each condition). Scale bar represents 5 μm. The graph shows the quantification of the TrkA/phalloidin staining at the plasma membrane over time in samples treated with NGF plus the indicated protein or buffer. (B) Mouse SCG dissociated neurons were grown during 7 DIV using microfluidic devices. Neurons were deprived of NGF for 16 h and NGF and viral proteins were added to the distal axon compartment during 120 min. TrkA phosphorylation was analyzed by immunofluorescence after permeabilization with Triton X-100 and normalized using phalloidin. The images and the graph correspond to the same experiment and are representative of three independent experiments. Scale bar 10 μm. n = 10 fields for each condition. A two-tailed unpaired T-test was used to calculate significance in (A) and (B) . *** P

    Article Snippet: In brief, 340 μL of rat tail collagen I (BD Biosciences, San Jose, CA) were mixed with 40 μL of 10x MEM, 10 μL of HEPES or the vCKBPs at a final concentration of 50 nM, and mouse NGF 2.5S (N-100, Alomone labs, Jerusalem, Israel) at a final concentration of 0.25 nM.

    Techniques: Immunofluorescence, Staining, Two Tailed Test

    SgG2 modifies NGF-TrkA signaling. Mouse SCG dissociated neurons were grown during 5 days in vitro (DIV). (A) Neurons were deprived of NGF for 16 h and were stimulated with NGF alone, NGF plus SgG2 or SgG2 alone during 5 min. TrkA-SgG2 interaction was analyzed by TrkA immunoprecipitation (IP) followed by Western blot (WB) to detect SgG2. Molecular sizes in kDa and the position of SgG2 (empty arrowhead) and TrkA (solid arrowhead) are indicated. The experiment shown is representative of three independent assays. (B-F) Neurons were deprived of NGF for 16 h and were stimulated with NGF and the indicated viral proteins during 0, 15 and 120 min. (B) The phosphorylation levels of TrkA, ERK, AKT and cofilin were analyzed by Western blot (WB) using specific antibodies. Detection of actin was used as a loading control. All blots correspond to the same experiment. Graphs show statistical analysis for (C) p-TrkA, (D) p-ERK, (E) pAKT and (F) p-cofilin levels. p-TrkA n = 4; p-ERK n = 4; p-AKT n = 4; p-cofilin n = 3. To calculate significance a two-tailed unpaired T-test was employed; * P

    Journal: PLoS Pathogens

    Article Title: Secreted Herpes Simplex Virus-2 Glycoprotein G Modifies NGF-TrkA Signaling to Attract Free Nerve Endings to the Site of Infection

    doi: 10.1371/journal.ppat.1004571

    Figure Lengend Snippet: SgG2 modifies NGF-TrkA signaling. Mouse SCG dissociated neurons were grown during 5 days in vitro (DIV). (A) Neurons were deprived of NGF for 16 h and were stimulated with NGF alone, NGF plus SgG2 or SgG2 alone during 5 min. TrkA-SgG2 interaction was analyzed by TrkA immunoprecipitation (IP) followed by Western blot (WB) to detect SgG2. Molecular sizes in kDa and the position of SgG2 (empty arrowhead) and TrkA (solid arrowhead) are indicated. The experiment shown is representative of three independent assays. (B-F) Neurons were deprived of NGF for 16 h and were stimulated with NGF and the indicated viral proteins during 0, 15 and 120 min. (B) The phosphorylation levels of TrkA, ERK, AKT and cofilin were analyzed by Western blot (WB) using specific antibodies. Detection of actin was used as a loading control. All blots correspond to the same experiment. Graphs show statistical analysis for (C) p-TrkA, (D) p-ERK, (E) pAKT and (F) p-cofilin levels. p-TrkA n = 4; p-ERK n = 4; p-AKT n = 4; p-cofilin n = 3. To calculate significance a two-tailed unpaired T-test was employed; * P

    Article Snippet: In brief, 340 μL of rat tail collagen I (BD Biosciences, San Jose, CA) were mixed with 40 μL of 10x MEM, 10 μL of HEPES or the vCKBPs at a final concentration of 50 nM, and mouse NGF 2.5S (N-100, Alomone labs, Jerusalem, Israel) at a final concentration of 0.25 nM.

    Techniques: In Vitro, Immunoprecipitation, Western Blot, Two Tailed Test

    SgG2 increases NGF-dependent axonal growth. (A,B,D) Mouse SCGs were grown as explants in collagen matrix in the presence of the indicated trophic factors and viral proteins. Neurons were stained with Tuj1 antibody targeting class III ß–tubulin. Nuclei were stained with TO-PRO-3. Images are a projection of at least 3 stacks, correspond to the same experiment, and are representative of three (A,D) or two (B) independent experiments. The graphs below represent the quantification from three (A,D) or two (B) independent experiments. To calculate significance, a two-tailed unpaired T-test was applied. *** P

    Journal: PLoS Pathogens

    Article Title: Secreted Herpes Simplex Virus-2 Glycoprotein G Modifies NGF-TrkA Signaling to Attract Free Nerve Endings to the Site of Infection

    doi: 10.1371/journal.ppat.1004571

    Figure Lengend Snippet: SgG2 increases NGF-dependent axonal growth. (A,B,D) Mouse SCGs were grown as explants in collagen matrix in the presence of the indicated trophic factors and viral proteins. Neurons were stained with Tuj1 antibody targeting class III ß–tubulin. Nuclei were stained with TO-PRO-3. Images are a projection of at least 3 stacks, correspond to the same experiment, and are representative of three (A,D) or two (B) independent experiments. The graphs below represent the quantification from three (A,D) or two (B) independent experiments. To calculate significance, a two-tailed unpaired T-test was applied. *** P

    Article Snippet: In brief, 340 μL of rat tail collagen I (BD Biosciences, San Jose, CA) were mixed with 40 μL of 10x MEM, 10 μL of HEPES or the vCKBPs at a final concentration of 50 nM, and mouse NGF 2.5S (N-100, Alomone labs, Jerusalem, Israel) at a final concentration of 0.25 nM.

    Techniques: Staining, Two Tailed Test

    SgG2 promotes the incorporation of TrkA into different microdomains of the plasma membrane. Mouse SCG dissociated neurons were grown during 5 DIV. Neurons were deprived of NGF for 16 h and subsequently stimulated with NGF, vCKBPs or both. Colocalization between TrkA and two different subtypes of lipid rafts was studied using immunofluorescence without permeabilization. TrkA colocalization with GM1 rafts 2 min (A) and 10 min (B) post-stimulation. TrkA colocalization with GM3 rafts 2 min (C) , and 10 min (D) post-stimulation. Confocal microscopy images correspond to one representative cell from each condition. The +ves image displays pseudocolored pixels from the areas within the plasma membrane in which both TrkA and the corresponding subtype of lipid rafts pixel value exceed the mean. Scale bar represents 5 μm. Graphs show the average values (mean±SEM) obtained for PC and ICQ. Plots in (A) represent n = 18–26 neurons from three independent assays. Plots in (B) show the results obtained for 15 cells from two independent experiments. Plots in (C) represent n = 21–35 neurons from three independent assays. Plots in (D) represent 16–23 neurons from two independent assays. Two-tailed unpaired T-test, * P

    Journal: PLoS Pathogens

    Article Title: Secreted Herpes Simplex Virus-2 Glycoprotein G Modifies NGF-TrkA Signaling to Attract Free Nerve Endings to the Site of Infection

    doi: 10.1371/journal.ppat.1004571

    Figure Lengend Snippet: SgG2 promotes the incorporation of TrkA into different microdomains of the plasma membrane. Mouse SCG dissociated neurons were grown during 5 DIV. Neurons were deprived of NGF for 16 h and subsequently stimulated with NGF, vCKBPs or both. Colocalization between TrkA and two different subtypes of lipid rafts was studied using immunofluorescence without permeabilization. TrkA colocalization with GM1 rafts 2 min (A) and 10 min (B) post-stimulation. TrkA colocalization with GM3 rafts 2 min (C) , and 10 min (D) post-stimulation. Confocal microscopy images correspond to one representative cell from each condition. The +ves image displays pseudocolored pixels from the areas within the plasma membrane in which both TrkA and the corresponding subtype of lipid rafts pixel value exceed the mean. Scale bar represents 5 μm. Graphs show the average values (mean±SEM) obtained for PC and ICQ. Plots in (A) represent n = 18–26 neurons from three independent assays. Plots in (B) show the results obtained for 15 cells from two independent experiments. Plots in (C) represent n = 21–35 neurons from three independent assays. Plots in (D) represent 16–23 neurons from two independent assays. Two-tailed unpaired T-test, * P

    Article Snippet: In brief, 340 μL of rat tail collagen I (BD Biosciences, San Jose, CA) were mixed with 40 μL of 10x MEM, 10 μL of HEPES or the vCKBPs at a final concentration of 50 nM, and mouse NGF 2.5S (N-100, Alomone labs, Jerusalem, Israel) at a final concentration of 0.25 nM.

    Techniques: Immunofluorescence, Confocal Microscopy, Two Tailed Test

    BDNF-induced NR2C upregulation by TrkB activation. Cells were cultured in low KCl and treated with or without BDNF (100 ng/ml in A and 50 ng/ml in B-D and F ) for 96 h. A , Levels of indicated mRNAs were quantitated by RNA blotting ( n = 4). B , Cell lysates (40 μg) were immunoblotted with anti-panNR2 antibody or anti-NR1 antibody. Molecular sizes (kilodaltons) of protein makers are indicated on the left. C , P2 membrane fractions were isolated, solubilized, and immunoprecipitated (IP) with anti-NR1 antibody, followed by immnoblotting with anti-panNR2 antibody. D , Cell-surface proteins were biotinylated with Sulfo-NHS-SS-Biotin. Cell lysates were solubilized, precipitated with NeutrAvidin beads, and immunoblotted with anti-panNR2 antibody. E , Cells were cultured in low KCl and treated with BDNF, NGF, NT-3, or NT-4 (50 ng/ml each) for 96 h. Levels of NR2C mRNA were quantitated ( n = 4). F , Granule cells were prepared from TrkB - / - knock-out mice(KO) and their wild-type (WT) littermates. NR2C and TrkB mRNAs were analyzed by RNA blotting. Ethidium bromide-stained 18s rRNA is also indicated. * p

    Journal: The Journal of Neuroscience

    Article Title: Neuronal Depolarization Controls Brain-Derived Neurotrophic Factor-Induced Upregulation of NR2C NMDA Receptor via Calcineurin Signaling

    doi: 10.1523/JNEUROSCI.2191-05.2005

    Figure Lengend Snippet: BDNF-induced NR2C upregulation by TrkB activation. Cells were cultured in low KCl and treated with or without BDNF (100 ng/ml in A and 50 ng/ml in B-D and F ) for 96 h. A , Levels of indicated mRNAs were quantitated by RNA blotting ( n = 4). B , Cell lysates (40 μg) were immunoblotted with anti-panNR2 antibody or anti-NR1 antibody. Molecular sizes (kilodaltons) of protein makers are indicated on the left. C , P2 membrane fractions were isolated, solubilized, and immunoprecipitated (IP) with anti-NR1 antibody, followed by immnoblotting with anti-panNR2 antibody. D , Cell-surface proteins were biotinylated with Sulfo-NHS-SS-Biotin. Cell lysates were solubilized, precipitated with NeutrAvidin beads, and immunoblotted with anti-panNR2 antibody. E , Cells were cultured in low KCl and treated with BDNF, NGF, NT-3, or NT-4 (50 ng/ml each) for 96 h. Levels of NR2C mRNA were quantitated ( n = 4). F , Granule cells were prepared from TrkB - / - knock-out mice(KO) and their wild-type (WT) littermates. NR2C and TrkB mRNAs were analyzed by RNA blotting. Ethidium bromide-stained 18s rRNA is also indicated. * p

    Article Snippet: Slices were treated with or without 1 μ m FK506 for 96 h, and 0.5 vol of the culture medium was changed every 2 d. Reagents and inhibitors were purchased from the following sources: BDNF (Peprotech, London, UK); neurotrophin-3 (NT-3), NT-4, and nerve growth factor (NGF) (Alomone Laboratories, Jerusalem, Israel); neuregulin-β (NeoMarkers, Fremont, CA); nifedipine (Nacalai Tesque, Kyoto, Japan); KN62 (Tocris, Bristol, UK); and U0126, SB203580, FK506, K252a, , PD98059, cyclosporin A, rapamycin, and bisindolylmaleimide I (Calbiochem, San Diego, CA).

    Techniques: Activation Assay, Cell Culture, Isolation, Immunoprecipitation, Knock-Out, Mouse Assay, Staining