α bungarotoxin  (Alomone Labs)


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  • 94
    Name:
    alpha Bungarotoxin
    Description:
    A Potent Antagonist of α7 and Muscle nAChR and GABA A Receptor Subtypes
    Catalog Number:
    B-100
    Price:
    127.0
    Category:
    Toxin
    Source:
    Natural peptide isolated from Bungarus multicinctus (Many-banded krait).
    Applications:
    0
    Purity:
    >99% (HPLC)
    Size:
    0 5 mg
    Format:
    Lyophilized powder.
    Formula:
    C338H529N97O105S11
    Molecular Weight:
    7984 Da.
    Molecule Name:
    alpha-bungarotoxin, Long neurotoxin 1, alpha-Bgtx, alpha-BuTX, alpha-BTX
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    Structured Review

    Alomone Labs α bungarotoxin
    alpha Bungarotoxin
    A Potent Antagonist of α7 and Muscle nAChR and GABA A Receptor Subtypes
    https://www.bioz.com/result/α bungarotoxin/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    α bungarotoxin - by Bioz Stars, 2021-09
    94/100 stars

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    Related Articles

    Software:

    Article Title: Differentiation of control and ALS mutant human iPSCs into functional skeletal muscle cells, a tool for the study of neuromuscolar diseases
    Article Snippet: .. ACh dose–response were studied applying to each cell three different doses of the transmitter (3, 30, 300 μM) for 1 s. ACh-evoked currents were blocked treating cells with α-bungarotoxin (5 μg/ml; α-Bungarotoxin-ATTO-488, Alomone Labs) for 2 h. Data were analyzed off line with Clampfit 10 sofware; Origin 7 software was used for statistical analysis of electrophysiological data. ..

    other:

    Article Title: Nicotinic Acetylcholine Receptors Containing the α7-Like Subunit Mediate Contractions of Muscles Responsible for Space Positioning of the Snail, Helix pomatia L. Tentacle
    Article Snippet: The α-bungarotoxin (αBgTx), α-conotoxin ImI (α-CTx IMI) and αA-conotoxin PIVA (αA-CTx PIVA) were purchased from Alomone Labs (Jerusalem, Israel).

    Imaging:

    Article Title: Defective Neuronal Positioning Correlates With Aberrant Motor Circuit Function in Zebrafish
    Article Snippet: .. One hour before imaging, larvae were injected with 4.6 ng of α-bungarotoxin (Alomone Labs) to paralyze and prevent twitching. ..

    Injection:

    Article Title: Defective Neuronal Positioning Correlates With Aberrant Motor Circuit Function in Zebrafish
    Article Snippet: .. One hour before imaging, larvae were injected with 4.6 ng of α-bungarotoxin (Alomone Labs) to paralyze and prevent twitching. ..

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  • 93
    Alomone Labs probdnf protein
    Schematic diagram showing how <t>proBDNF</t> dampens CD4 + T cell activity and contributes to the pathogenesis of SAE. In sepsis, proBDNF expression is increased in peripheral blood and meningeal immune cells, which then decreases the infiltration of CD4 + T cells in the meninges. As a result, meningeal pro-inflammatory cytokines such as IL-6 and IL-1β are upregulated, but anti-inflammatory cytokines including IL-4 and IL-13 are downregulated, finally leading to SAE. SAE, sepsis-associated encephalopathy
    Probdnf Protein, 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
    https://www.bioz.com/result/probdnf protein/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    probdnf protein - by Bioz Stars, 2021-09
    93/100 stars
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    93
    Alomone Labs recombinant human bdnf protein
    Rearing larvae in dim light reduces production of <t>BDNF</t> in the medial PGZ by 10 <t>dpf</t> ( A–C ). Scale bar, 15 μm. Injecting 5 dpf larvae with 100 ng/ml recombinant human BDNF increased the amount of BDNF protein detected by immunohistochemistry
    Recombinant Human Bdnf Protein, 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
    https://www.bioz.com/result/recombinant human bdnf protein/product/Alomone Labs
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    recombinant human bdnf protein - by Bioz Stars, 2021-09
    93/100 stars
      Buy from Supplier

    94
    Alomone Labs kv3 1b
    <t>Kv3.1b</t> is a component of the PNS nodes in Trembler-J nerves. These are images of unfixed teased sciatic nerve fibers from WT and Trembler-J ( TrJ ) mice, labeled as indicated. In WT mice, a few nodes are Kv3.1b positive (apposed arrowheads in A ), whereas
    Kv3 1b, 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/kv3 1b/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    kv3 1b - by Bioz Stars, 2021-09
    94/100 stars
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    94
    Alomone Labs forskolin
    CREB phosphorylation mediates AKT2-regulated induction of PEPCK. ( A ) CREB phosphorylation induced by PTEN loss is regulated by AKT2. Liver lysates from mice with different genotypes were analysis for p-CREB. PM, Pten deleted; DM, Pten and Akt2 deleted; AM, Akt2 deleted. Two biological replicas are included for each genotype group. Protein levels of p-AKT, PTEN and p-GSK3 are analyzed and showed up-regulation of AKT and GSK-3 with PTEN loss and inhibition when AKT2 is lost. Phosphorylation of CREB is only observed when PTEN is lost. AKT2 loss together with PTEN blocked this phosphorylation in the DM livers. ( B ) Akt2 +/+ and Akt2 − / − cell were treated with <t>forskolin</t> (100 μM) for 24 h to induce cAMP and CREB phosphorylation or vehicle as controls. Phosphorylation of CREB is detected and showed to be lower when AKT2 is lost. When forskolin is added to induce cAMP, protein expression of p-CREB is evaluated in both Akt2 +/+ and Akt2 − / − cells even though its levels remains lower in the Akt2 − / − cells. Vinculin is probed as loading control. ( C ) Introduction of CREB-133A and wtCREB to Akt2 +/+ and Akt2 −/− hepatocytes. Expression of PEPCK is analyzed using quantitative PCR analysis. Left panel, CREB-S133A is introduced to Akt2 +/+ cells. Middle panel, wtCREB is introduced to Akt2 −/− cells; Right panel, wtCREB is introduced to Akt2 +/+ cells. n = 3. Data presented as mean ± SEM. * P ≤ 0.05. Different from vector transfected cells of the same genotype. Experiments repeated at least three times.
    Forskolin, 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/forskolin/product/Alomone Labs
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    forskolin - by Bioz Stars, 2021-09
    94/100 stars
      Buy from Supplier

    Image Search Results


    Schematic diagram showing how proBDNF dampens CD4 + T cell activity and contributes to the pathogenesis of SAE. In sepsis, proBDNF expression is increased in peripheral blood and meningeal immune cells, which then decreases the infiltration of CD4 + T cells in the meninges. As a result, meningeal pro-inflammatory cytokines such as IL-6 and IL-1β are upregulated, but anti-inflammatory cytokines including IL-4 and IL-13 are downregulated, finally leading to SAE. SAE, sepsis-associated encephalopathy

    Journal: Journal of Neuroinflammation

    Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

    doi: 10.1186/s12974-020-01850-0

    Figure Lengend Snippet: Schematic diagram showing how proBDNF dampens CD4 + T cell activity and contributes to the pathogenesis of SAE. In sepsis, proBDNF expression is increased in peripheral blood and meningeal immune cells, which then decreases the infiltration of CD4 + T cells in the meninges. As a result, meningeal pro-inflammatory cytokines such as IL-6 and IL-1β are upregulated, but anti-inflammatory cytokines including IL-4 and IL-13 are downregulated, finally leading to SAE. SAE, sepsis-associated encephalopathy

    Article Snippet: 4 × 105 cells were put in each well of a 96-well flat-bottom plate and stimulated with 100, 200, or 500 ng ml−1 proBDNF protein (Alomone Labs, Israel, catalog: B243) as introduced by our previous studies [ ], respectively.

    Techniques: Activity Assay, Expressing

    Exogenous proBDNF protein reduced CD4 + T cells but increased CD8 + T cells of septic mice in vitro. Mice injected with saline or LPS (5 mg kg −1 ) for 5 days and the splenocytes were isolated and cultured for 3 days in vitro. Exogenous proBDNF did not alter the percentage of a CD3 + T cells in CD45 + cells or the percentage of b CD4 + T cells or c CD8 + T cells in CD3 + T cells in splenocytes from mice treated with saline. d Exogenous proBDNF did not alter the percentage of CD3 + T cells in CD45 + cells in splenocytes in septic mice. e–f ProBDNF treatment significantly decreased the percentage of e CD4 + T cells but increased the percentage of f CD8 + T cells in CD3 + T cells in splenocytes in LPS-treated mice. n = 4 in each group. Data were analyzed by one-way ANOVA and followed by Tukey post hoc test, * P

    Journal: Journal of Neuroinflammation

    Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

    doi: 10.1186/s12974-020-01850-0

    Figure Lengend Snippet: Exogenous proBDNF protein reduced CD4 + T cells but increased CD8 + T cells of septic mice in vitro. Mice injected with saline or LPS (5 mg kg −1 ) for 5 days and the splenocytes were isolated and cultured for 3 days in vitro. Exogenous proBDNF did not alter the percentage of a CD3 + T cells in CD45 + cells or the percentage of b CD4 + T cells or c CD8 + T cells in CD3 + T cells in splenocytes from mice treated with saline. d Exogenous proBDNF did not alter the percentage of CD3 + T cells in CD45 + cells in splenocytes in septic mice. e–f ProBDNF treatment significantly decreased the percentage of e CD4 + T cells but increased the percentage of f CD8 + T cells in CD3 + T cells in splenocytes in LPS-treated mice. n = 4 in each group. Data were analyzed by one-way ANOVA and followed by Tukey post hoc test, * P

    Article Snippet: 4 × 105 cells were put in each well of a 96-well flat-bottom plate and stimulated with 100, 200, or 500 ng ml−1 proBDNF protein (Alomone Labs, Israel, catalog: B243) as introduced by our previous studies [ ], respectively.

    Techniques: Mouse Assay, In Vitro, Injection, Isolation, Cell Culture

    Increased proBDNF expression in meningeal and peripheral blood immune cells in septic mice. Mice were i.p. injected with LPS (5 mg kg −1 ) and meninges and peripheral blood were harvested for immunofluorescence staining or flow cytometry. a Representative whole mount meningeal immunofluorescence images showed markedly increased proBDNF-positive staining cells in the meninges in mice at 1 day after LPS injection compared to saline injected mice. The high magnification images around the arrows are displayed in a white square as insets. Bar = 100 μm. b – f Representative meningeal single cell flow cytometry images ( upper panel ) and its statistical analysis ( lower panel ) indicated that proBDNF MFI was increased in meningeal b CD3 + T cells, c CD4 + T cells, d CD8 + T cells, and f CD11b + monocytes/macrophages at 1 day after LPS injection. proBDNF in meningeal e CD19 + B cells upregulated until 5 days after LPS injection. n = 10 in the Con group, n = 6 in LPS groups. g – k Upregulation of proBDNF in g CD3 + T cells, h CD4 + T cells, i CD8 + T cells, j CD19 + B cells, and k CD11b + monocytes/macrophages in peripheral blood in LPS-injected mice were detected. n = 9 in Con group, n = 8 in the LPS1d group, n = 4 in the LPS 5d group. Data b – k were analyzed by one-way ANOVA and followed by Tukey post hoc test, * P

    Journal: Journal of Neuroinflammation

    Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

    doi: 10.1186/s12974-020-01850-0

    Figure Lengend Snippet: Increased proBDNF expression in meningeal and peripheral blood immune cells in septic mice. Mice were i.p. injected with LPS (5 mg kg −1 ) and meninges and peripheral blood were harvested for immunofluorescence staining or flow cytometry. a Representative whole mount meningeal immunofluorescence images showed markedly increased proBDNF-positive staining cells in the meninges in mice at 1 day after LPS injection compared to saline injected mice. The high magnification images around the arrows are displayed in a white square as insets. Bar = 100 μm. b – f Representative meningeal single cell flow cytometry images ( upper panel ) and its statistical analysis ( lower panel ) indicated that proBDNF MFI was increased in meningeal b CD3 + T cells, c CD4 + T cells, d CD8 + T cells, and f CD11b + monocytes/macrophages at 1 day after LPS injection. proBDNF in meningeal e CD19 + B cells upregulated until 5 days after LPS injection. n = 10 in the Con group, n = 6 in LPS groups. g – k Upregulation of proBDNF in g CD3 + T cells, h CD4 + T cells, i CD8 + T cells, j CD19 + B cells, and k CD11b + monocytes/macrophages in peripheral blood in LPS-injected mice were detected. n = 9 in Con group, n = 8 in the LPS1d group, n = 4 in the LPS 5d group. Data b – k were analyzed by one-way ANOVA and followed by Tukey post hoc test, * P

    Article Snippet: 4 × 105 cells were put in each well of a 96-well flat-bottom plate and stimulated with 100, 200, or 500 ng ml−1 proBDNF protein (Alomone Labs, Israel, catalog: B243) as introduced by our previous studies [ ], respectively.

    Techniques: Expressing, Mouse Assay, Injection, Immunofluorescence, Staining, Flow Cytometry

    Effect of i.c.v. injection of anti-proBDNF antibody on fear memory and meningeal immune activity in the septic mice. Mice were bilateral i.c.v. injected with 1 μg McAb-proB 3 days before LPS injection. Behavior tests were performed 1 day after LPS injection. a Representative images showed the broad and thorough diffusion of drugs in cerebroventricular lumen following i.c.v. injection with methylene blue. b McAb-proB i.c.v. injection did not influence weight of mice. c–e There was no difference of c fear conditioning acquiring performance in each group, nor McAb-proB increased the freezing time of d contextual or e cued fear conditioning test as compared to IgG control after LPS injection. n = 6 in each group. Data b , c , and e were analyzed by repeated measures ANOVA and followed by Bonferroni post hoc test and data d was analyzed by one-way ANOVA and followed by Tukey post hoc test, * P

    Journal: Journal of Neuroinflammation

    Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

    doi: 10.1186/s12974-020-01850-0

    Figure Lengend Snippet: Effect of i.c.v. injection of anti-proBDNF antibody on fear memory and meningeal immune activity in the septic mice. Mice were bilateral i.c.v. injected with 1 μg McAb-proB 3 days before LPS injection. Behavior tests were performed 1 day after LPS injection. a Representative images showed the broad and thorough diffusion of drugs in cerebroventricular lumen following i.c.v. injection with methylene blue. b McAb-proB i.c.v. injection did not influence weight of mice. c–e There was no difference of c fear conditioning acquiring performance in each group, nor McAb-proB increased the freezing time of d contextual or e cued fear conditioning test as compared to IgG control after LPS injection. n = 6 in each group. Data b , c , and e were analyzed by repeated measures ANOVA and followed by Bonferroni post hoc test and data d was analyzed by one-way ANOVA and followed by Tukey post hoc test, * P

    Article Snippet: 4 × 105 cells were put in each well of a 96-well flat-bottom plate and stimulated with 100, 200, or 500 ng ml−1 proBDNF protein (Alomone Labs, Israel, catalog: B243) as introduced by our previous studies [ ], respectively.

    Techniques: Injection, Activity Assay, Mouse Assay, Diffusion-based Assay

    Systemic blockade of proBDNF ameliorated cognitive dysfunction and restored meningeal and peripheral CD4 + T cell ratio in septic mice. Mice were i.p. injected with proBDNF 30 min before LPS (5 mg kg −1 ) injection. Fear conditioning testing was performed 1 day after LPS injection. Meninges and peripheral blood were harvested 5 days after LPS injection for flow cytometry. a McAb-proB did not influence the weight of mice or b fear conditioning acquiring. c , d McAb-proB greatly alleviated memory deficit induced by LPS injection in mice as indicated by the increased freezing time in ( c ) contextual and ( d ) cued fear conditioning tests in the McAb-proB group relative to the IgG control. n = 8 in each group. Data a , b , and d were analyzed by repeated measures ANOVA and followed by Bonferroni post hoc tests. Data c was analyzed by one-way ANOVA and followed by Tukey post hoc test, * P

    Journal: Journal of Neuroinflammation

    Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

    doi: 10.1186/s12974-020-01850-0

    Figure Lengend Snippet: Systemic blockade of proBDNF ameliorated cognitive dysfunction and restored meningeal and peripheral CD4 + T cell ratio in septic mice. Mice were i.p. injected with proBDNF 30 min before LPS (5 mg kg −1 ) injection. Fear conditioning testing was performed 1 day after LPS injection. Meninges and peripheral blood were harvested 5 days after LPS injection for flow cytometry. a McAb-proB did not influence the weight of mice or b fear conditioning acquiring. c , d McAb-proB greatly alleviated memory deficit induced by LPS injection in mice as indicated by the increased freezing time in ( c ) contextual and ( d ) cued fear conditioning tests in the McAb-proB group relative to the IgG control. n = 8 in each group. Data a , b , and d were analyzed by repeated measures ANOVA and followed by Bonferroni post hoc tests. Data c was analyzed by one-way ANOVA and followed by Tukey post hoc test, * P

    Article Snippet: 4 × 105 cells were put in each well of a 96-well flat-bottom plate and stimulated with 100, 200, or 500 ng ml−1 proBDNF protein (Alomone Labs, Israel, catalog: B243) as introduced by our previous studies [ ], respectively.

    Techniques: Mouse Assay, Injection, Flow Cytometry

    Systemic blockade of proBDNF restored meningeal pro-inflammatory microenvironment in septic mice. Mice were i.p. injected with proBDNF 30 min before LPS (5 mg kg −1 ) injection. Meninges were harvested 5 days after LPS injection for qPCR. a The level of CD4 gene expression was higher in the meninges of the McAb-proB group than in IgG controls in septic mice. b–f Gene levels were significantly lower in b IL-1β and c IL-6 but higher in d IL-4, e IFN-γ, and f IL-13 in the meninges after LPS injection in the McAb-proB group as compared to IgG control. n = 5 in each group. All experiments were performed at least in triplicate. Data were analyzed by unpaired T test, * P

    Journal: Journal of Neuroinflammation

    Article Title: ProBDNF promotes sepsis-associated encephalopathy in mice by dampening the immune activity of meningeal CD4+ T cells

    doi: 10.1186/s12974-020-01850-0

    Figure Lengend Snippet: Systemic blockade of proBDNF restored meningeal pro-inflammatory microenvironment in septic mice. Mice were i.p. injected with proBDNF 30 min before LPS (5 mg kg −1 ) injection. Meninges were harvested 5 days after LPS injection for qPCR. a The level of CD4 gene expression was higher in the meninges of the McAb-proB group than in IgG controls in septic mice. b–f Gene levels were significantly lower in b IL-1β and c IL-6 but higher in d IL-4, e IFN-γ, and f IL-13 in the meninges after LPS injection in the McAb-proB group as compared to IgG control. n = 5 in each group. All experiments were performed at least in triplicate. Data were analyzed by unpaired T test, * P

    Article Snippet: 4 × 105 cells were put in each well of a 96-well flat-bottom plate and stimulated with 100, 200, or 500 ng ml−1 proBDNF protein (Alomone Labs, Israel, catalog: B243) as introduced by our previous studies [ ], respectively.

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

    Rearing larvae in dim light reduces production of BDNF in the medial PGZ by 10 dpf ( A–C ). Scale bar, 15 μm. Injecting 5 dpf larvae with 100 ng/ml recombinant human BDNF increased the amount of BDNF protein detected by immunohistochemistry

    Journal: The Journal of Neuroscience

    Article Title: Visual Experience Facilitates BDNF-Dependent Adaptive Recruitment of New Neurons in the Postembryonic Optic Tectum

    doi: 10.1523/JNEUROSCI.1962-17.2018

    Figure Lengend Snippet: Rearing larvae in dim light reduces production of BDNF in the medial PGZ by 10 dpf ( A–C ). Scale bar, 15 μm. Injecting 5 dpf larvae with 100 ng/ml recombinant human BDNF increased the amount of BDNF protein detected by immunohistochemistry

    Article Snippet: To test whether exogenous BDNF was detectable in whole-brain lysates following ICV injections, we performed a Western blot using 5 dpf larvae injected with either 100 ng/ml recombinant human BDNF protein (#B-250, Alomone Labs) or a vehicle control.

    Techniques: Recombinant, Immunohistochemistry

    Kv3.1b is a component of the PNS nodes in Trembler-J nerves. These are images of unfixed teased sciatic nerve fibers from WT and Trembler-J ( TrJ ) mice, labeled as indicated. In WT mice, a few nodes are Kv3.1b positive (apposed arrowheads in A ), whereas

    Journal: The Journal of Neuroscience

    Article Title: Altered Ion Channels in an Animal Model of Charcot-Marie-Tooth Disease Type IA

    doi: 10.1523/JNEUROSCI.3328-04.2005

    Figure Lengend Snippet: Kv3.1b is a component of the PNS nodes in Trembler-J nerves. These are images of unfixed teased sciatic nerve fibers from WT and Trembler-J ( TrJ ) mice, labeled as indicated. In WT mice, a few nodes are Kv3.1b positive (apposed arrowheads in A ), whereas

    Article Snippet: Frozen sections and teased fibers were permeabilized by immersion in -20°C acetone for 10 min, blocked at room temperature for 1 h with 5% fish skin gelatin containing 0.1% Triton X-100 in PBS, and incubated overnight at 4°C with various combinations of primary antibodies: rabbit antisera against KCNQ2 [1:200 ( )], Kv3.1b (1:100; Alomone Labs, Jerusalem, Israel), Kv1.2 (1:100; Alomone Labs), Nav1.6 [1:100 ( )], Nav1.8 [1:100 ( )], Nav1.8 [1:500 ( )], ankyrin-G [1:100 ( ; )], contactin [1:100 ( )], ezrin-binding protein 50 kDa (EBP-50) (ab3452; 1:100; Abcam, Cambridge, MA), syndecan-3 [1:300 ( )], or Caspr [1:500 ( )]; and mouse monoclonal antibodies against panNav channels (K58/35; 1:250; Sigma, St. Louis, MO), Nav1.2 [1:100 ( ; )], Nav1.2 (1:50; Upstate Biotechnology, Lake Placid, NY), Caspr [1:50 ( )], annexin II light chain (1:50; BD Transduction Laboratories, San Jose, CA), or myelin-associated glycoprotein (MAG) (clone 513, 1:100; Boehringer Mannheim, Indianapolis, IN); rat monoclonal antibody against E-cadherin (1:50; Sigma); chicken antibody against βIV-spectrin [1:500 ( )]; and a soluble construct of receptor protein tyrosine phosphatase β conjugated to the human Fc fragment (RPTPβ-Fc) [1:3 ( )].

    Techniques: Mouse Assay, Labeling

    CREB phosphorylation mediates AKT2-regulated induction of PEPCK. ( A ) CREB phosphorylation induced by PTEN loss is regulated by AKT2. Liver lysates from mice with different genotypes were analysis for p-CREB. PM, Pten deleted; DM, Pten and Akt2 deleted; AM, Akt2 deleted. Two biological replicas are included for each genotype group. Protein levels of p-AKT, PTEN and p-GSK3 are analyzed and showed up-regulation of AKT and GSK-3 with PTEN loss and inhibition when AKT2 is lost. Phosphorylation of CREB is only observed when PTEN is lost. AKT2 loss together with PTEN blocked this phosphorylation in the DM livers. ( B ) Akt2 +/+ and Akt2 − / − cell were treated with forskolin (100 μM) for 24 h to induce cAMP and CREB phosphorylation or vehicle as controls. Phosphorylation of CREB is detected and showed to be lower when AKT2 is lost. When forskolin is added to induce cAMP, protein expression of p-CREB is evaluated in both Akt2 +/+ and Akt2 − / − cells even though its levels remains lower in the Akt2 − / − cells. Vinculin is probed as loading control. ( C ) Introduction of CREB-133A and wtCREB to Akt2 +/+ and Akt2 −/− hepatocytes. Expression of PEPCK is analyzed using quantitative PCR analysis. Left panel, CREB-S133A is introduced to Akt2 +/+ cells. Middle panel, wtCREB is introduced to Akt2 −/− cells; Right panel, wtCREB is introduced to Akt2 +/+ cells. n = 3. Data presented as mean ± SEM. * P ≤ 0.05. Different from vector transfected cells of the same genotype. Experiments repeated at least three times.

    Journal: The Biochemical journal

    Article Title: Regulation of basal expression of hepatic PEPCK and G6Pase by AKT2

    doi: 10.1042/BCJ20190570

    Figure Lengend Snippet: CREB phosphorylation mediates AKT2-regulated induction of PEPCK. ( A ) CREB phosphorylation induced by PTEN loss is regulated by AKT2. Liver lysates from mice with different genotypes were analysis for p-CREB. PM, Pten deleted; DM, Pten and Akt2 deleted; AM, Akt2 deleted. Two biological replicas are included for each genotype group. Protein levels of p-AKT, PTEN and p-GSK3 are analyzed and showed up-regulation of AKT and GSK-3 with PTEN loss and inhibition when AKT2 is lost. Phosphorylation of CREB is only observed when PTEN is lost. AKT2 loss together with PTEN blocked this phosphorylation in the DM livers. ( B ) Akt2 +/+ and Akt2 − / − cell were treated with forskolin (100 μM) for 24 h to induce cAMP and CREB phosphorylation or vehicle as controls. Phosphorylation of CREB is detected and showed to be lower when AKT2 is lost. When forskolin is added to induce cAMP, protein expression of p-CREB is evaluated in both Akt2 +/+ and Akt2 − / − cells even though its levels remains lower in the Akt2 − / − cells. Vinculin is probed as loading control. ( C ) Introduction of CREB-133A and wtCREB to Akt2 +/+ and Akt2 −/− hepatocytes. Expression of PEPCK is analyzed using quantitative PCR analysis. Left panel, CREB-S133A is introduced to Akt2 +/+ cells. Middle panel, wtCREB is introduced to Akt2 −/− cells; Right panel, wtCREB is introduced to Akt2 +/+ cells. n = 3. Data presented as mean ± SEM. * P ≤ 0.05. Different from vector transfected cells of the same genotype. Experiments repeated at least three times.

    Article Snippet: The medium was replaced with fresh medium with 100 μM Forskolin (Alomone Labs Cat# F-500) or vehicle and incubated for 24 h before harvesting.

    Techniques: Mouse Assay, Inhibition, Expressing, Real-time Polymerase Chain Reaction, Plasmid Preparation, Transfection