trichostatin a  (Alomone Labs)


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  • 85
    Name:
    Trichostatin A
    Description:
    Trichostatin A an antifungal antibiotic irreversibly inhibits histone deacetylase It could also promote apoptosis and in some cases even inhibits the proliferation of some cancer cells
    Catalog Number:
    T-300
    Price:
    554.0
    Category:
    Small Molecule
    Source:
    Streptomyces platensis.
    Applications:
    0
    Purity:
    >98%
    Size:
    1 Vials containing 1 mg each
    Format:
    Lyophilized/solid.
    Formula:
    C17H22N2O3
    Molecular Weight:
    302.4
    Molecule Name:
    (2E,4E,6R)-7-(4-(Dimethylamino)phenyl)-N-hydroxy-4,6- di methyl-7-oxo-2,4-heptadienamide.
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    Structured Review

    Alomone Labs trichostatin a
    Trichostatin A
    Trichostatin A an antifungal antibiotic irreversibly inhibits histone deacetylase It could also promote apoptosis and in some cases even inhibits the proliferation of some cancer cells
    https://www.bioz.com/result/trichostatin a/product/Alomone Labs
    Average 85 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    trichostatin a - by Bioz Stars, 2021-09
    85/100 stars

    Images

    1) Product Images from "IL12-Mediated Liver Inflammation Reduces the Formation of AAV Transcriptionally Active Forms but Has No Effect over Preexisting AAV Transgene Expression"

    Article Title: IL12-Mediated Liver Inflammation Reduces the Formation of AAV Transcriptionally Active Forms but Has No Effect over Preexisting AAV Transgene Expression

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0067748

    DNA demethylating agents but not histone acetylation inhibition revert the IFN-γ inhibitory effect. C57BL/6 mice received 2.5 × 1011 vg/kg of AAV8-luc vector alone or in combination with 1.5 × 109 vg/kg of AAV8-IL12 vector. One group of 5 mice treated with AAV8-luc or AAV8-luc in combination with AAV8-IL12 received 5′-Azacytidine (AZA) intraperitoneally at a dose of 1 mg/kg every 24 hours starting the day of vector injection. A second group of 5 mice treated with AAV8-luc or AAV8-luc in combination with AAV8-IL12 received 2 mg/kg of trichostatin A (TSA) were injected IP every 48 hours starting the day of vector injection. A third group of animals received only vector injection. Luciferase expression levels were analyzed in both groups of animals at days 1, 4, and 7 using a Xenogen in vivo luminometer, experiments could not be performed at longer time points due to the toxicity of 5-AZA long term treatment. Luciferase expression was quantified and represented as photons/second. The data are shown as mean ± standard deviation (SD). The differences in luciferase expression were statistically evaluated by Student t test (**p
    Figure Legend Snippet: DNA demethylating agents but not histone acetylation inhibition revert the IFN-γ inhibitory effect. C57BL/6 mice received 2.5 × 1011 vg/kg of AAV8-luc vector alone or in combination with 1.5 × 109 vg/kg of AAV8-IL12 vector. One group of 5 mice treated with AAV8-luc or AAV8-luc in combination with AAV8-IL12 received 5′-Azacytidine (AZA) intraperitoneally at a dose of 1 mg/kg every 24 hours starting the day of vector injection. A second group of 5 mice treated with AAV8-luc or AAV8-luc in combination with AAV8-IL12 received 2 mg/kg of trichostatin A (TSA) were injected IP every 48 hours starting the day of vector injection. A third group of animals received only vector injection. Luciferase expression levels were analyzed in both groups of animals at days 1, 4, and 7 using a Xenogen in vivo luminometer, experiments could not be performed at longer time points due to the toxicity of 5-AZA long term treatment. Luciferase expression was quantified and represented as photons/second. The data are shown as mean ± standard deviation (SD). The differences in luciferase expression were statistically evaluated by Student t test (**p

    Techniques Used: Inhibition, Mouse Assay, Plasmid Preparation, Injection, Luciferase, Expressing, In Vivo, Standard Deviation

    Related Articles

    Inhibition:

    Article Title: IL12-Mediated Liver Inflammation Reduces the Formation of AAV Transcriptionally Active Forms but Has No Effect over Preexisting AAV Transgene Expression
    Article Snippet: .. For the inhibition of histone deacetylation, 2 mg/kg of trichostatin A (Alomon Laboratories) were injected IP every 48 hours. ..

    Injection:

    Article Title: IL12-Mediated Liver Inflammation Reduces the Formation of AAV Transcriptionally Active Forms but Has No Effect over Preexisting AAV Transgene Expression
    Article Snippet: .. For the inhibition of histone deacetylation, 2 mg/kg of trichostatin A (Alomon Laboratories) were injected IP every 48 hours. ..

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  • 95
    Alomone Labs rabbit α hcn2 antibody
    Domains mediating the interaction of VAPB with <t>HCN2.</t> A ) Schematic illustration of a HCN subunit. The CNBD and some of the truncation constructs studied are indicated. B ) All truncation constructs exhibited a positive interaction, evident from growth on -LWHA dropout medium. C ) Representative current traces and the relative currents for different C-terminal deletions expressed alone or with VAPB. D ) Representative current traces and the relative current amplitudes for the N-terminal truncated NTK HCN2 expressed alone or with VAPB. E ) Relative current amplitudes of NTK HCN2 HA Ex (extracellular HA-tag) expressed alone or with VAPB. F ) Relative surface expression of NTK HCN2 HA Ex expressed alone or with VAPB analyzed as relative light units (RLUs). G ) Schematic illustration, representative traces, and currents of a HCN2 channel chimera with the N terminus of HCN4 ( HCN4-N HCN2) expressed alone or with VAPB. H ) Relative currents of HCN2 expressed alone or coexpressed with VAPB (1.7 ± 0.1), TM VAPB (1.6 ± 0.2), the MSP domain (MSP VAPB ), the MSP with half of the CC domain (MSP-CC 0.5 VAPB ), or with the complete CC domain (MSP-CC VAPB ). I , J ) Relative current amplitudes of HCN2 HA Ex expressed alone or with TM VAPB (1.3 ± 0.1) ( I ) and the respective changes in the relative surface membrane expression analyzed as RLUs, using a single cell chemiluminescence assay (TM VAPB 1.8 ± 0.2) ( J ). All data are presented as means ± sem . The number of experiments ( n ) is indicated in the respective bar graphs. N.s., not significant. * P
    Rabbit α Hcn2 Antibody, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit α hcn2 antibody/product/Alomone Labs
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    95
    Alomone Labs ω conotoxin gvia
    Rb1 inhibited the I Ba in hippocampal neurons, and this inhibitory effect was eliminated after the application of nifedipine (A). Neither <t>ω-conotoxin-GVIA</t> nor ω-agatoxin IVA diminished the Rb1-sensitive I Ba (B and C, respectively). Upper panel, pairs of the inward currents evoked by pulses from −60 to +0 mV (0–+20 mV) at the times indicated in the lower panel. Lower panel, time course of the effects of 10 μmol/L Rb1 on the I Ba amplitude before and after application of the Ca 2+ channel antagonists (10 μmol/L nifedipine, 1 μmol/L ω-conotoxin GVIA and 30 nmol/L ω-agatoxin IVA). The bar graphs for Rb1 inhibition (mean±SEM, n =5 for Rb1) on the I Ba in cells untreated or treated with Ca 2+ channel antagonists. c P
    ω Conotoxin Gvia, supplied by Alomone Labs, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Alomone Labs anti aqp4
    The synthesized GILZ-p inhibited LPS induced Müller cell gliosis. Western blot analysis was performed to determine the protein expression levels of glial fibrillary acidic protein (GFAP) (A,B) , and <t>AQP4</t> (C,D) in Müller cells treated with 1000 ng/ml LPS in combination with different concentrations of GILZ-p (0.01, 0.1, 1, and 10 μM) for 24 h. β-actin was used as the loading control. The results of quantitative analysis, as determined by densitometric analysis, were expressed as relative to β-actin. Data represent the mean ± SE; the Mann–Whitney U -test was used for comparisons between two groups. n = 3 for each group. ∗ P
    Anti Aqp4, 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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    Image Search Results


    Domains mediating the interaction of VAPB with HCN2. A ) Schematic illustration of a HCN subunit. The CNBD and some of the truncation constructs studied are indicated. B ) All truncation constructs exhibited a positive interaction, evident from growth on -LWHA dropout medium. C ) Representative current traces and the relative currents for different C-terminal deletions expressed alone or with VAPB. D ) Representative current traces and the relative current amplitudes for the N-terminal truncated NTK HCN2 expressed alone or with VAPB. E ) Relative current amplitudes of NTK HCN2 HA Ex (extracellular HA-tag) expressed alone or with VAPB. F ) Relative surface expression of NTK HCN2 HA Ex expressed alone or with VAPB analyzed as relative light units (RLUs). G ) Schematic illustration, representative traces, and currents of a HCN2 channel chimera with the N terminus of HCN4 ( HCN4-N HCN2) expressed alone or with VAPB. H ) Relative currents of HCN2 expressed alone or coexpressed with VAPB (1.7 ± 0.1), TM VAPB (1.6 ± 0.2), the MSP domain (MSP VAPB ), the MSP with half of the CC domain (MSP-CC 0.5 VAPB ), or with the complete CC domain (MSP-CC VAPB ). I , J ) Relative current amplitudes of HCN2 HA Ex expressed alone or with TM VAPB (1.3 ± 0.1) ( I ) and the respective changes in the relative surface membrane expression analyzed as RLUs, using a single cell chemiluminescence assay (TM VAPB 1.8 ± 0.2) ( J ). All data are presented as means ± sem . The number of experiments ( n ) is indicated in the respective bar graphs. N.s., not significant. * P

    Journal: The FASEB Journal

    Article Title: The VAMP-associated protein VAPB is required for cardiac and neuronal pacemaker channel function

    doi: 10.1096/fj.201800246R

    Figure Lengend Snippet: Domains mediating the interaction of VAPB with HCN2. A ) Schematic illustration of a HCN subunit. The CNBD and some of the truncation constructs studied are indicated. B ) All truncation constructs exhibited a positive interaction, evident from growth on -LWHA dropout medium. C ) Representative current traces and the relative currents for different C-terminal deletions expressed alone or with VAPB. D ) Representative current traces and the relative current amplitudes for the N-terminal truncated NTK HCN2 expressed alone or with VAPB. E ) Relative current amplitudes of NTK HCN2 HA Ex (extracellular HA-tag) expressed alone or with VAPB. F ) Relative surface expression of NTK HCN2 HA Ex expressed alone or with VAPB analyzed as relative light units (RLUs). G ) Schematic illustration, representative traces, and currents of a HCN2 channel chimera with the N terminus of HCN4 ( HCN4-N HCN2) expressed alone or with VAPB. H ) Relative currents of HCN2 expressed alone or coexpressed with VAPB (1.7 ± 0.1), TM VAPB (1.6 ± 0.2), the MSP domain (MSP VAPB ), the MSP with half of the CC domain (MSP-CC 0.5 VAPB ), or with the complete CC domain (MSP-CC VAPB ). I , J ) Relative current amplitudes of HCN2 HA Ex expressed alone or with TM VAPB (1.3 ± 0.1) ( I ) and the respective changes in the relative surface membrane expression analyzed as RLUs, using a single cell chemiluminescence assay (TM VAPB 1.8 ± 0.2) ( J ). All data are presented as means ± sem . The number of experiments ( n ) is indicated in the respective bar graphs. N.s., not significant. * P

    Article Snippet: Untagged HCN2 protein was detected with rabbit α-HCN2 antibody (APC-030, 1:300; Alomone Labs, Jerusalem, Israel) and peroxidase-conjugated goat α-rabbit IgG antibody (32460, 1:2000; Thermo Fisher Scientific) as the secondary antibody.

    Techniques: Construct, Expressing, Chemiluminescence Immunoassay

    VAPB determines surface expression and dendritic localization of HCN2. A ) Live cell imaging of HeLa cells transfected with an N-terminally EGFP-tagged HCN2 carrying an extracellular HA-epitope ( EGFP HCN2 HA Ex ) alone or cotransfected with VAPB or the TM segment of VAPB (TM VAPB ). B ) Chemiluminescence assays of fixed non-permeabilized HeLa cells, analyzing the surface expression as relative light units (RLUs) for EGFP HCN2 HA Ex alone and after cotransfection with VAPB (1.6 ± 0.1). Upper inset illustrates a representative control Western blot showing an unaltered HCN2 prote in expression. C ) Chemiluminescence surface expression assay as in B , but using TM VAPB (1.6 ± 0.1). D ) Immunocytochemistry of HA VAPB transfected cortical neurons. Endogenous HCN2 (green) is colocalizing (white) with HA VAPB (magenta) in the soma and dendrites. Anti–MAP2-staining illustrating an intact neuronal network and dendrites (blue). E ) Immunocytochemistry experiment as in D , but transfecting the ALS8 mutation HA VAPB P56S (magenta), leading to an aggregation of VAPB P56S in the soma of the neurons. Also, HCN2 fluorescence (green) was focused in the soma and dendritic localization was lost, despite an intact neuronal network (α-MAP2, blue). Scale bars, 20 µm ( A , D , E ). All data are presented as means ± sem . The number of experiments ( n ) is indicated in the respective bar graphs. ** P

    Journal: The FASEB Journal

    Article Title: The VAMP-associated protein VAPB is required for cardiac and neuronal pacemaker channel function

    doi: 10.1096/fj.201800246R

    Figure Lengend Snippet: VAPB determines surface expression and dendritic localization of HCN2. A ) Live cell imaging of HeLa cells transfected with an N-terminally EGFP-tagged HCN2 carrying an extracellular HA-epitope ( EGFP HCN2 HA Ex ) alone or cotransfected with VAPB or the TM segment of VAPB (TM VAPB ). B ) Chemiluminescence assays of fixed non-permeabilized HeLa cells, analyzing the surface expression as relative light units (RLUs) for EGFP HCN2 HA Ex alone and after cotransfection with VAPB (1.6 ± 0.1). Upper inset illustrates a representative control Western blot showing an unaltered HCN2 prote in expression. C ) Chemiluminescence surface expression assay as in B , but using TM VAPB (1.6 ± 0.1). D ) Immunocytochemistry of HA VAPB transfected cortical neurons. Endogenous HCN2 (green) is colocalizing (white) with HA VAPB (magenta) in the soma and dendrites. Anti–MAP2-staining illustrating an intact neuronal network and dendrites (blue). E ) Immunocytochemistry experiment as in D , but transfecting the ALS8 mutation HA VAPB P56S (magenta), leading to an aggregation of VAPB P56S in the soma of the neurons. Also, HCN2 fluorescence (green) was focused in the soma and dendritic localization was lost, despite an intact neuronal network (α-MAP2, blue). Scale bars, 20 µm ( A , D , E ). All data are presented as means ± sem . The number of experiments ( n ) is indicated in the respective bar graphs. ** P

    Article Snippet: Untagged HCN2 protein was detected with rabbit α-HCN2 antibody (APC-030, 1:300; Alomone Labs, Jerusalem, Israel) and peroxidase-conjugated goat α-rabbit IgG antibody (32460, 1:2000; Thermo Fisher Scientific) as the secondary antibody.

    Techniques: Expressing, Live Cell Imaging, Transfection, Cotransfection, Western Blot, Immunocytochemistry, Staining, Mutagenesis, Fluorescence

    Codistribution of VAPs with HCN2 and contribution to thalamic I h . A – E ), Distribution of HCN2, VAPB, and VAPA mRNA in mouse brain and spinal cord. ISH analysis of HCN2, VAPB, and VAPA using DIG-labeled riboprobes, revealing mRNA expression of VAPB in cortical areas ( A ), hippocampus ( B ), thalamus ( C ), cerebellum ( D ) (arrows point to interneurons in the granular layer), and spinal cord ( E ). Note the overlapping distribution of VAPB with HCN2 and VAPA mRNA. Am, amygdala; CA, cornu ammonis; DG, dentate gyrus; DH, dorsal horn; gcl, granule cell layer; Hb, habenulae; ic, internal capsule; LG, lateral geniculate ncl.; m, molecular cell layer; pcl, Purkinje cell layer; RTh, reticular thalamic ncl.; Sth, subthalamic ncl.; VB, ventrobasal thalamus; Th, thalamus; VH, ventral horn. F ) Representative current traces elicited in slice patch-clamp experiments of the ventrobasal thalamus (VB) of wild-type animals (control) and VAPB −/− mice. G ) The I h current was significantly reduced in VAPB −/− mice (15.4 ± 1.1 pA/pF) compared with control animals (22.2 ± 2.3 pA/pF). H ) Average activation curves of the VB I h current for control and VAPB −/− mice. V 1/2 of activation for control (−91.6 ± 1.3 mV, n = 8) and VAPB −/− (−87.5 ± 1.2 mV, n = 7). Scale bars: 500 µm ( A–C , E ), 100 µm ( D ). All data are presented as means ± sem . The number of experiments ( n ) is indicated in the respective bar graphs. * P

    Journal: The FASEB Journal

    Article Title: The VAMP-associated protein VAPB is required for cardiac and neuronal pacemaker channel function

    doi: 10.1096/fj.201800246R

    Figure Lengend Snippet: Codistribution of VAPs with HCN2 and contribution to thalamic I h . A – E ), Distribution of HCN2, VAPB, and VAPA mRNA in mouse brain and spinal cord. ISH analysis of HCN2, VAPB, and VAPA using DIG-labeled riboprobes, revealing mRNA expression of VAPB in cortical areas ( A ), hippocampus ( B ), thalamus ( C ), cerebellum ( D ) (arrows point to interneurons in the granular layer), and spinal cord ( E ). Note the overlapping distribution of VAPB with HCN2 and VAPA mRNA. Am, amygdala; CA, cornu ammonis; DG, dentate gyrus; DH, dorsal horn; gcl, granule cell layer; Hb, habenulae; ic, internal capsule; LG, lateral geniculate ncl.; m, molecular cell layer; pcl, Purkinje cell layer; RTh, reticular thalamic ncl.; Sth, subthalamic ncl.; VB, ventrobasal thalamus; Th, thalamus; VH, ventral horn. F ) Representative current traces elicited in slice patch-clamp experiments of the ventrobasal thalamus (VB) of wild-type animals (control) and VAPB −/− mice. G ) The I h current was significantly reduced in VAPB −/− mice (15.4 ± 1.1 pA/pF) compared with control animals (22.2 ± 2.3 pA/pF). H ) Average activation curves of the VB I h current for control and VAPB −/− mice. V 1/2 of activation for control (−91.6 ± 1.3 mV, n = 8) and VAPB −/− (−87.5 ± 1.2 mV, n = 7). Scale bars: 500 µm ( A–C , E ), 100 µm ( D ). All data are presented as means ± sem . The number of experiments ( n ) is indicated in the respective bar graphs. * P

    Article Snippet: Untagged HCN2 protein was detected with rabbit α-HCN2 antibody (APC-030, 1:300; Alomone Labs, Jerusalem, Israel) and peroxidase-conjugated goat α-rabbit IgG antibody (32460, 1:2000; Thermo Fisher Scientific) as the secondary antibody.

    Techniques: In Situ Hybridization, Labeling, Expressing, Patch Clamp, Mouse Assay, Activation Assay

    VAPB selectively increases HCN1 and HCN2 currents. A ) Y2H direct interaction assay. Transformation control (-LW), leucine, and tryptophan dropout. Interaction read-out (-LWHA), additional dropout of histidine and adenine. pAL-Alg5, positive control. pPR3-N, negative control. B ) Topology of VAPB. C ) GST VAPB pull-down of HCN2 EGFP using transfected HeLa cells. D ) GST VAPA, GST VAMP1, or GST VAMP2 pull-down of HCN2 EGFP using transfected HeLa cells. E ) GST VAPA pull-down of HCN2 and endogenous VAPB, using HCN2 EGFP transfected HeLa cells. F ) Pull-down of in vitro translated HCN2 (untagged). G ) Pull-down of HCN2 from rat brain lysates. H , I ) Representative currents ( H ) of HCN2 expressed in oocytes alone or with VAPB and the relative current amplitudes ( I ) analyzed over 3 d. J ) Relative currents of HCN1, HCN2, and HCN4 alone or coexpressed with VAPB. K , L ) Relative currents of different potassium channels ( K ) coexpressed with VAPB and of HCN2 ( L ) coexpressed with VAPA, VAPB, or VAPC. M ) Relative currents of HCN2 coexpressed with a mixture of VAPA/B (1:1). N ) Representative macropatch recordings in different configurations: on cell (o.c.), inside-out after patch excision (i.o.), and after application of 100 µM cAMP (i.o.+100 µM cAMP). O , P ) Activation curves for HCN2 alone ( n = 6) ( O ), recorded as in N , or after coexpression with VAPB ( n = 8) ( P ). Q ) V 1/2 values for HCN2 expressed alone or with VAPB in different patch modes. R ) Relative currents of HCN2 HA Ex alone or with VAPB. S ) Relative surface expression of HCN2 HA Ex expressed alone or with VAPB, analyzed as relative light units (RLUs). T ) Relative currents of HCN2 expressed alone or with VAPB or VAPB P56S . All data are presented as means ± sem . The number of cells ( n ) is indicated in the bar graphs. N.s., not significant. * P

    Journal: The FASEB Journal

    Article Title: The VAMP-associated protein VAPB is required for cardiac and neuronal pacemaker channel function

    doi: 10.1096/fj.201800246R

    Figure Lengend Snippet: VAPB selectively increases HCN1 and HCN2 currents. A ) Y2H direct interaction assay. Transformation control (-LW), leucine, and tryptophan dropout. Interaction read-out (-LWHA), additional dropout of histidine and adenine. pAL-Alg5, positive control. pPR3-N, negative control. B ) Topology of VAPB. C ) GST VAPB pull-down of HCN2 EGFP using transfected HeLa cells. D ) GST VAPA, GST VAMP1, or GST VAMP2 pull-down of HCN2 EGFP using transfected HeLa cells. E ) GST VAPA pull-down of HCN2 and endogenous VAPB, using HCN2 EGFP transfected HeLa cells. F ) Pull-down of in vitro translated HCN2 (untagged). G ) Pull-down of HCN2 from rat brain lysates. H , I ) Representative currents ( H ) of HCN2 expressed in oocytes alone or with VAPB and the relative current amplitudes ( I ) analyzed over 3 d. J ) Relative currents of HCN1, HCN2, and HCN4 alone or coexpressed with VAPB. K , L ) Relative currents of different potassium channels ( K ) coexpressed with VAPB and of HCN2 ( L ) coexpressed with VAPA, VAPB, or VAPC. M ) Relative currents of HCN2 coexpressed with a mixture of VAPA/B (1:1). N ) Representative macropatch recordings in different configurations: on cell (o.c.), inside-out after patch excision (i.o.), and after application of 100 µM cAMP (i.o.+100 µM cAMP). O , P ) Activation curves for HCN2 alone ( n = 6) ( O ), recorded as in N , or after coexpression with VAPB ( n = 8) ( P ). Q ) V 1/2 values for HCN2 expressed alone or with VAPB in different patch modes. R ) Relative currents of HCN2 HA Ex alone or with VAPB. S ) Relative surface expression of HCN2 HA Ex expressed alone or with VAPB, analyzed as relative light units (RLUs). T ) Relative currents of HCN2 expressed alone or with VAPB or VAPB P56S . All data are presented as means ± sem . The number of cells ( n ) is indicated in the bar graphs. N.s., not significant. * P

    Article Snippet: Untagged HCN2 protein was detected with rabbit α-HCN2 antibody (APC-030, 1:300; Alomone Labs, Jerusalem, Israel) and peroxidase-conjugated goat α-rabbit IgG antibody (32460, 1:2000; Thermo Fisher Scientific) as the secondary antibody.

    Techniques: Transformation Assay, Positive Control, Negative Control, Transfection, In Vitro, Activation Assay, Expressing

    Rb1 inhibited the I Ba in hippocampal neurons, and this inhibitory effect was eliminated after the application of nifedipine (A). Neither ω-conotoxin-GVIA nor ω-agatoxin IVA diminished the Rb1-sensitive I Ba (B and C, respectively). Upper panel, pairs of the inward currents evoked by pulses from −60 to +0 mV (0–+20 mV) at the times indicated in the lower panel. Lower panel, time course of the effects of 10 μmol/L Rb1 on the I Ba amplitude before and after application of the Ca 2+ channel antagonists (10 μmol/L nifedipine, 1 μmol/L ω-conotoxin GVIA and 30 nmol/L ω-agatoxin IVA). The bar graphs for Rb1 inhibition (mean±SEM, n =5 for Rb1) on the I Ba in cells untreated or treated with Ca 2+ channel antagonists. c P

    Journal: Acta Pharmacologica Sinica

    Article Title: Ginsenoside Rb1 selectively inhibits the activity of L-type voltage-gated calcium channels in cultured rat hippocampal neurons

    doi: 10.1038/aps.2011.181

    Figure Lengend Snippet: Rb1 inhibited the I Ba in hippocampal neurons, and this inhibitory effect was eliminated after the application of nifedipine (A). Neither ω-conotoxin-GVIA nor ω-agatoxin IVA diminished the Rb1-sensitive I Ba (B and C, respectively). Upper panel, pairs of the inward currents evoked by pulses from −60 to +0 mV (0–+20 mV) at the times indicated in the lower panel. Lower panel, time course of the effects of 10 μmol/L Rb1 on the I Ba amplitude before and after application of the Ca 2+ channel antagonists (10 μmol/L nifedipine, 1 μmol/L ω-conotoxin GVIA and 30 nmol/L ω-agatoxin IVA). The bar graphs for Rb1 inhibition (mean±SEM, n =5 for Rb1) on the I Ba in cells untreated or treated with Ca 2+ channel antagonists. c P

    Article Snippet: Stock solutions of the Ca2+ channel antagonists, ω-conotoxin GVIA (Alomone Labs, UK), nifedipine (Sigma, UK) ω-agatoxin IVA (Alomone Labs, UK) and adenylyl cyclase agonist Forskolin (Sigma, UK), were prepared with the appropriate amounts of deionized water or dimethyl sulfoxide (DMSO) and frozen at −20 °C before appropriate dilution in the recording medium.

    Techniques: Inhibition

    (A) Phase-contrast image showing a single patch recording from 7-d cultured hippocampal neurons for the recording of the VGCCs. Scale bar, 10 μm. (B) Pharmacological separation of the VGCC subtypes in hippocampal neurons. Upper panel, inward Ca 2+ channel Ba 2+ currents evoked by pulses from −60 mV to 0 mV at the times indicated in the lower panel. Lower panel, time course of effects of ω-conotoxin GVIA (1 μmol/L), ω-agatoxin IVA (30 nmol/L) and nifedipine (10 μmol/L) on the Ba 2+ current amplitude.

    Journal: Acta Pharmacologica Sinica

    Article Title: Ginsenoside Rb1 selectively inhibits the activity of L-type voltage-gated calcium channels in cultured rat hippocampal neurons

    doi: 10.1038/aps.2011.181

    Figure Lengend Snippet: (A) Phase-contrast image showing a single patch recording from 7-d cultured hippocampal neurons for the recording of the VGCCs. Scale bar, 10 μm. (B) Pharmacological separation of the VGCC subtypes in hippocampal neurons. Upper panel, inward Ca 2+ channel Ba 2+ currents evoked by pulses from −60 mV to 0 mV at the times indicated in the lower panel. Lower panel, time course of effects of ω-conotoxin GVIA (1 μmol/L), ω-agatoxin IVA (30 nmol/L) and nifedipine (10 μmol/L) on the Ba 2+ current amplitude.

    Article Snippet: Stock solutions of the Ca2+ channel antagonists, ω-conotoxin GVIA (Alomone Labs, UK), nifedipine (Sigma, UK) ω-agatoxin IVA (Alomone Labs, UK) and adenylyl cyclase agonist Forskolin (Sigma, UK), were prepared with the appropriate amounts of deionized water or dimethyl sulfoxide (DMSO) and frozen at −20 °C before appropriate dilution in the recording medium.

    Techniques: Cell Culture

    The effect of L-type voltage-dependent Ca 2+ channels (VDCCs) on branching morphogenesis. ( A ) Morphological changes of SMG cultures (E13.5) upon 500 μM LaCl 3 treatment. ( B ) Bud numbers of SMG cultures upon 500 μM LaCl 3 (La) and 1 M EGTA treatment. n = 7, Data are represented as mean ± SEM. ( C ) Representative images of SMG cultures treated with various Ca 2+ channel inhibitors. ( D ) Bud numbers of SMG cultures (E12) upon treatment with various Ca 2+ channel inhibitors. Nif: 100 μM nifedipine; Gd: 500 μM GdCl3; SKF: 10 μM SKF 96365, n = 7, Data are represented as mean ±SEM. ( E ) Bud numbers of SMG cultures (E13) upon different concentrations of nifedipine treatment for 48 h. n = 5. Data are represented as mean ± SEM. ( F ) Relative acinar size of SMGs (E13) upon different concentrations of nifedipine treatment. n = 5. Data are represented as mean ±SEM. ( G ) Epithelial bud numbers of SMGs (E13.5) upon treatment with antagonists for different types of VDCCs: 2 μM w-Agatoxin IVA (Aga, P-type); 2 μM SNX 482 (SNX, R-type); 10 μM w-Conotoxin GVIA (Cono, N-type). n = 6. Data are represented as mean ±SEM. ( H ) Time-course changes of bud outline of developing SMG cultures. Arrowheads indicate the cleft initiation points. ( I ) Time-lapse images of epithelial rudiment cultures (E13) upon 100 μM nifedipine treatment. Arrowheads indicate cleft sites. Panels below indicate the bud numbers. Scale bars: 200 ( A , C ), 100 μm ( H , I ).

    Journal: Scientific Reports

    Article Title: Voltage-dependent Ca2+ channels promote branching morphogenesis of salivary glands by patterning differential growth

    doi: 10.1038/s41598-018-25957-w

    Figure Lengend Snippet: The effect of L-type voltage-dependent Ca 2+ channels (VDCCs) on branching morphogenesis. ( A ) Morphological changes of SMG cultures (E13.5) upon 500 μM LaCl 3 treatment. ( B ) Bud numbers of SMG cultures upon 500 μM LaCl 3 (La) and 1 M EGTA treatment. n = 7, Data are represented as mean ± SEM. ( C ) Representative images of SMG cultures treated with various Ca 2+ channel inhibitors. ( D ) Bud numbers of SMG cultures (E12) upon treatment with various Ca 2+ channel inhibitors. Nif: 100 μM nifedipine; Gd: 500 μM GdCl3; SKF: 10 μM SKF 96365, n = 7, Data are represented as mean ±SEM. ( E ) Bud numbers of SMG cultures (E13) upon different concentrations of nifedipine treatment for 48 h. n = 5. Data are represented as mean ± SEM. ( F ) Relative acinar size of SMGs (E13) upon different concentrations of nifedipine treatment. n = 5. Data are represented as mean ±SEM. ( G ) Epithelial bud numbers of SMGs (E13.5) upon treatment with antagonists for different types of VDCCs: 2 μM w-Agatoxin IVA (Aga, P-type); 2 μM SNX 482 (SNX, R-type); 10 μM w-Conotoxin GVIA (Cono, N-type). n = 6. Data are represented as mean ±SEM. ( H ) Time-course changes of bud outline of developing SMG cultures. Arrowheads indicate the cleft initiation points. ( I ) Time-lapse images of epithelial rudiment cultures (E13) upon 100 μM nifedipine treatment. Arrowheads indicate cleft sites. Panels below indicate the bud numbers. Scale bars: 200 ( A , C ), 100 μm ( H , I ).

    Article Snippet: The chemical reagents used in this study are as follows: 100 μM nifedipine (Sigma-Aldrich, St. Louis, MO; N7634); 500 μM gadolinium chloride (Sigma-Aldrich, G7532); 10 μM (Sigma-Aldrich, S7809); 1 M EGTA (Sigma-Aldrich, E4378); 500 μM lanthanum chloride (Sigma-Aldrich, L4131); 2 μM ω-Agatoxin IVA (Tocris, Bristol, UK; 2799); 2 μM SNX 482 (Tocris, 2799); 10 μM ω-Conotoxin GVIA (Alomone Labs, C-300); 10 μM U0126 (Sigma-Aldrich, U120); 50 mM potassium chloride (Sigma-Aldrich, P3911); 100 nM AP24534 (Tocris, 4274); 25 μM trifluoperazine dihydrochloride (Sigma Aldrich, T8516).

    Techniques:

    The synthesized GILZ-p inhibited LPS induced Müller cell gliosis. Western blot analysis was performed to determine the protein expression levels of glial fibrillary acidic protein (GFAP) (A,B) , and AQP4 (C,D) in Müller cells treated with 1000 ng/ml LPS in combination with different concentrations of GILZ-p (0.01, 0.1, 1, and 10 μM) for 24 h. β-actin was used as the loading control. The results of quantitative analysis, as determined by densitometric analysis, were expressed as relative to β-actin. Data represent the mean ± SE; the Mann–Whitney U -test was used for comparisons between two groups. n = 3 for each group. ∗ P

    Journal: Frontiers in Pharmacology

    Article Title: A Synthesized Glucocorticoid- Induced Leucine Zipper Peptide Inhibits Retinal Müller Cell Gliosis

    doi: 10.3389/fphar.2018.00331

    Figure Lengend Snippet: The synthesized GILZ-p inhibited LPS induced Müller cell gliosis. Western blot analysis was performed to determine the protein expression levels of glial fibrillary acidic protein (GFAP) (A,B) , and AQP4 (C,D) in Müller cells treated with 1000 ng/ml LPS in combination with different concentrations of GILZ-p (0.01, 0.1, 1, and 10 μM) for 24 h. β-actin was used as the loading control. The results of quantitative analysis, as determined by densitometric analysis, were expressed as relative to β-actin. Data represent the mean ± SE; the Mann–Whitney U -test was used for comparisons between two groups. n = 3 for each group. ∗ P

    Article Snippet: The membranes were blocked in 5% non-fat milk at room temperature for 1 h and incubated with the following antibodies: anti-monocyte chemoattractant protein (MCP)-1 (ab25124; Abcam, Cambridge, United Kingdom), anti-intercellular adhesion molecule (ICAM)-1 (ab171123; Proteintech, Chicago, IL, United States), anti-IL1β (ab9787; Abcam), Anti-tumor necrosis factor (TNF)α (PB0270, Boster Biological Technology, Wuhan, China), rabbit anti-p65 polyclonal antibody (ab16502; Abcam), anti-phospho-NF-κB p65 (Ser536) rabbit monoclonal antibody (3033P; Cell Signaling Technology, Beverly, MA, United States), anti-AQP4 (300-314, Alomone Labs, Jerusalem, Israel), anti-glial fibrillary acidic protein (GFAP) (ab10062, Abcam), or rabbit anti β-actin antibody (ab69512; Abcam) overnight.

    Techniques: Synthesized, Western Blot, Expressing, MANN-WHITNEY