alpha-sarcoglycan Search Results


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  • 89
    Novocastra α sarcoglycan
    Restoration of dystrophin-associated proteins. ( Left ) Muscle of control mdx mouse. ( Right ) Muscle of mdx mouse after triple tail-vein injections of the 2OMeAO at weekly intervals. Membrane localized <t>α-sarcoglycan,</t> β-dystroglycan and neuronal nitric oxide synthase (nNOS) are detected in the fibers expressing dystrophin after the 2OMeAO treatment. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (blue).
    α Sarcoglycan, supplied by Novocastra, used in various techniques. Bioz Stars score: 89/100, based on 187 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novocastra β sarcoglycan
    Functional and phenotypic correction in mdx mice following treatments with PNA AOs at 100 mg/kg for five weekly intravenous injections . ( a ) Serial immunostaining of dystrophin-associated protein complex (DAPC) in mdx mice. DAPC protein components β-dystroglycan, <t>α-sarcoglycan,</t> and <t>β-sarcoglycan</t> and neuronal nitric oxide synthase (nNOS) were detected in quadriceps from mdx mice treated with PNA AOs. Arrowheads point to the identical muscle fibers (bar = 200 μm). ( b ) Muscle function was assessed using grip strength test to determine the physical improvement of PNA AOs treated mdx mice. Significant force recovery was detected in treated mdx mice compared with untreated mdx controls ( t -test, * P
    β Sarcoglycan, supplied by Novocastra, used in various techniques. Bioz Stars score: 89/100, based on 160 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novocastra γ sarcoglycan
    HDAC4 directly represses structural gene expression via binding and inhibition of MEF2 activity. A ) Promoter sequence of <t>γ-sarcoglycan</t> spanning −812 to −861 nt upstream of transcriptional start. AT-rich MEF2 sites are indicated above, which were mutated singly in or combination for subsequent luciferase assays. B ) C2C12 myoblasts were tranfected with luciferase reporter constructs containing wild-type or MEF2 binding site deletion (ΔAT1, ΔAT2, or ΔAT1,2) promoters as indicated. Transfections occurred in the presence of MEF2 and/or HDAC4 as indicated. Luciferase activity was determined 48 h after transfection. C . D ) ChIP analysis was performed on C2C12 myotubes using either control IgG-, MEF2-, or HDAC4-specific antibodies. Bound DNA was analyzed by standard PCR generating a 300 bp fragment encompassing both MEF2 binding sites in the γ-sarcoglycan promoter. Input lane represents 5% of total starting material.
    γ Sarcoglycan, supplied by Novocastra, used in various techniques. Bioz Stars score: 89/100, based on 89 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Novocastra δ sarcoglycan
    Generation of <t>dystrophin/δ-sarcoglycan</t> double mutant mice. ( A ) The breeding scheme. ( B ) Representative PCR genotyping results. Bkg, background; δSG KO, δ-sarcoglycan deficient mice; δ-Dko, δ-sarcoglycan/dystrophin
    δ Sarcoglycan, supplied by Novocastra, used in various techniques. Bioz Stars score: 89/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Novocastra adhalin
    The muscle biopsy of the patient with the genetically confirmed LGMD2D showing no sign of dystrophy (A) and normal expression of all <t>sarcoglycans</t> (B, C, D, E) . The biopsy was taken at the age of 8 years.
    Adhalin, supplied by Novocastra, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Takeda takeda s epsilon sarcoglycan
    The muscle biopsy of the patient with the genetically confirmed LGMD2D showing no sign of dystrophy (A) and normal expression of all <t>sarcoglycans</t> (B, C, D, E) . The biopsy was taken at the age of 8 years.
    Takeda S Epsilon Sarcoglycan, supplied by Takeda, used in various techniques. Bioz Stars score: 85/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Leica Biosystems α sarcoglycan
    Disruption of dystrophin expression in the skeletal muscle of DMD KO rabbits. (A-C) Immunofluorescence staining of muscle sections from WT and DMD KO rabbits with mouse monoclonal antibodies against dystrophin (A), glycosylated α-dystroglycan (B) and <t>α-sarcoglycan</t> (C). Nuclei were stained by DAPI. Scale bar: 100 µm.
    α Sarcoglycan, supplied by Leica Biosystems, used in various techniques. Bioz Stars score: 92/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Abcam δ sarcoglycan
    Tsp expression in muscle of Drosophila rescues MD due to deletion of the <t>δ-sarcoglycan-like</t> gene (Sgcd 840 ). ( A ) Fly survival was compared over a period of 40 days. p
    δ Sarcoglycan, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology δ sarcoglycan
    Generation of <t>dystrophin/δ-sarcoglycan</t> double mutant mice. ( A ) The breeding scheme. ( B ) Representative PCR genotyping results. Bkg, background; δSG KO, δ-sarcoglycan deficient mice; δ-Dko, δ-sarcoglycan/dystrophin
    δ Sarcoglycan, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 89/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Developmental Studies Hybridoma Bank α sarcoglycan
    Overexpression of Serpina3n restores the abundance of sarcolemma adhesion components in two dystrophic mouse models. ( A ) Immunohistochemistry of transverse quadriceps histological sections or ( B ) western blotting from quadriceps muscle of membrane protein extracts from mice at 2–3 months of age. The genotypes of mice used are shown with the indicated antibodies. Nuclei are stained in blue with DAPI, and Ponceau was used as a western loading control. Dystr. dystrophin; Utr, utrophin; β1D-Itg, β1D-integrin; α7-Itg, α7-integrin; δ-SG, <t>δ-sarcoglycan;</t> α-DG, α-dystroglycan; β-DG, β-dystroglycan; β-SG, β-sarcoglycan; α-SG; <t>α-sarcoglycan;</t> CaV1.1, L-type calcium channel. ( C ) Quantitative protein analysis for the indicated proteins in dystrophic muscle following overexpression of Serpina3n. A total of three separate animals for each genotype were used for densitometric analysis. * P
    α Sarcoglycan, supplied by Developmental Studies Hybridoma Bank, used in various techniques. Bioz Stars score: 90/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Leica Microsystems β sarcoglycan
    Differences in α-dystrobrevin splice variants between cardiac and skeletal muscle DAPC. ( A ) Western blot analysis of α-dystrobrevins in mouse cardiac (C) and skeletal (S) muscle total protein lysates, DYS-IPs and IgG-IPs. α3-dystrobrevin associates with dystrophin in the heart. Fold differences in α-dystrobrevin abundance in cardiac vs. skeletal muscle DYS-IPs relative to dystrophin are shown (averages ±SD, N = 3). ( B ) Immunolabeling of wild type cardiac sections for α1 and α2-dystrobrevins. Scale bar: 50 µm. ( C ) Immunolabeling of mdx cardiac tissue section for <t>β-sarcoglycan.</t> Scale bar: 50 µm. ( D ) Western blot analysis of α-dystrobrevins in heart protein lysates from wild type (WT) and mdx mice. Fold differences in α-dystrobrevin abundance in WT vs. mdx cardiac lysates relative to GAPDH are shown (averages ±SD, N = 3) ( E ) Western blot analysis of α-dystrobrevins in DYS-IPs from human (H) and mouse (M) cardiac samples. Additional α-dystrobrevin isoforms (arrow heads) are detected in human cardiac lysates and DYS-IP.
    β Sarcoglycan, supplied by Leica Microsystems, used in various techniques. Bioz Stars score: 89/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Leica Microsystems α sarcoglycan
    Differences in α-dystrobrevin splice variants between cardiac and skeletal muscle DAPC. ( A ) Western blot analysis of α-dystrobrevins in mouse cardiac (C) and skeletal (S) muscle total protein lysates, DYS-IPs and IgG-IPs. α3-dystrobrevin associates with dystrophin in the heart. Fold differences in α-dystrobrevin abundance in cardiac vs. skeletal muscle DYS-IPs relative to dystrophin are shown (averages ±SD, N = 3). ( B ) Immunolabeling of wild type cardiac sections for α1 and α2-dystrobrevins. Scale bar: 50 µm. ( C ) Immunolabeling of mdx cardiac tissue section for <t>β-sarcoglycan.</t> Scale bar: 50 µm. ( D ) Western blot analysis of α-dystrobrevins in heart protein lysates from wild type (WT) and mdx mice. Fold differences in α-dystrobrevin abundance in WT vs. mdx cardiac lysates relative to GAPDH are shown (averages ±SD, N = 3) ( E ) Western blot analysis of α-dystrobrevins in DYS-IPs from human (H) and mouse (M) cardiac samples. Additional α-dystrobrevin isoforms (arrow heads) are detected in human cardiac lysates and DYS-IP.
    α Sarcoglycan, supplied by Leica Microsystems, used in various techniques. Bioz Stars score: 89/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Novocastra mouse monoclonal anti α sarcoglycan
    Dependence of ecto-nucleotidase activity of <t>α-sarcoglycan-expressing</t> HEK-293 cells on the bivalent cation concentrations ( A ) ATP-hydrolysing activity was measured in stably transfected cells in the presence of 4 mM Mg 2+ or 2 mM Ca 2+ or both (4 mM Mg 2+ and 2 mM Ca 2+ ). ( B ) ATP-hydrolysing activity of α-sarcoglycan stably transfected HEK-293 cells was measured either in the presence of 4 mM Mg 2+ and the indicated concentrations of Ca 2+ (○) or in the presence of 2 mM Ca 2+ and the indicated concentrations of Mg 2+ (•). The plot shows the values after the subtraction of the empty vector-transfected activity. Results are from four experiments performed in triplicate. ** P
    Mouse Monoclonal Anti α Sarcoglycan, supplied by Novocastra, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology β sarcoglycan
    Dependence of ecto-nucleotidase activity of <t>α-sarcoglycan-expressing</t> HEK-293 cells on the bivalent cation concentrations ( A ) ATP-hydrolysing activity was measured in stably transfected cells in the presence of 4 mM Mg 2+ or 2 mM Ca 2+ or both (4 mM Mg 2+ and 2 mM Ca 2+ ). ( B ) ATP-hydrolysing activity of α-sarcoglycan stably transfected HEK-293 cells was measured either in the presence of 4 mM Mg 2+ and the indicated concentrations of Ca 2+ (○) or in the presence of 2 mM Ca 2+ and the indicated concentrations of Mg 2+ (•). The plot shows the values after the subtraction of the empty vector-transfected activity. Results are from four experiments performed in triplicate. ** P
    β Sarcoglycan, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 89/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Leica Biosystems γ sarcoglycan
    Dependence of ecto-nucleotidase activity of <t>α-sarcoglycan-expressing</t> HEK-293 cells on the bivalent cation concentrations ( A ) ATP-hydrolysing activity was measured in stably transfected cells in the presence of 4 mM Mg 2+ or 2 mM Ca 2+ or both (4 mM Mg 2+ and 2 mM Ca 2+ ). ( B ) ATP-hydrolysing activity of α-sarcoglycan stably transfected HEK-293 cells was measured either in the presence of 4 mM Mg 2+ and the indicated concentrations of Ca 2+ (○) or in the presence of 2 mM Ca 2+ and the indicated concentrations of Mg 2+ (•). The plot shows the values after the subtraction of the empty vector-transfected activity. Results are from four experiments performed in triplicate. ** P
    γ Sarcoglycan, supplied by Leica Biosystems, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Leica Microsystems γ sarcoglycan
    Dependence of ecto-nucleotidase activity of <t>α-sarcoglycan-expressing</t> HEK-293 cells on the bivalent cation concentrations ( A ) ATP-hydrolysing activity was measured in stably transfected cells in the presence of 4 mM Mg 2+ or 2 mM Ca 2+ or both (4 mM Mg 2+ and 2 mM Ca 2+ ). ( B ) ATP-hydrolysing activity of α-sarcoglycan stably transfected HEK-293 cells was measured either in the presence of 4 mM Mg 2+ and the indicated concentrations of Ca 2+ (○) or in the presence of 2 mM Ca 2+ and the indicated concentrations of Mg 2+ (•). The plot shows the values after the subtraction of the empty vector-transfected activity. Results are from four experiments performed in triplicate. ** P
    γ Sarcoglycan, supplied by Leica Microsystems, used in various techniques. Bioz Stars score: 89/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Santa Cruz Biotechnology γ sarcoglycan
    The expression analyses of α-, β-, <t>γ-sarcoglycan</t> and dysferlin protein in the muscles of patient by Western blot. The total proteins extracted from skeletal muscles of normal person (lane 1), relative normal muscle biopsies of patient IV-7 (lane 2), and dystrophic muscle biopsies of patient IV-7 (lane 3).
    γ Sarcoglycan, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 89/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Leica Biosystems beta sarcoglycan
    The expression analyses of α-, β-, <t>γ-sarcoglycan</t> and dysferlin protein in the muscles of patient by Western blot. The total proteins extracted from skeletal muscles of normal person (lane 1), relative normal muscle biopsies of patient IV-7 (lane 2), and dystrophic muscle biopsies of patient IV-7 (lane 3).
    Beta Sarcoglycan, supplied by Leica Biosystems, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Restoration of dystrophin-associated proteins. ( Left ) Muscle of control mdx mouse. ( Right ) Muscle of mdx mouse after triple tail-vein injections of the 2OMeAO at weekly intervals. Membrane localized α-sarcoglycan, β-dystroglycan and neuronal nitric oxide synthase (nNOS) are detected in the fibers expressing dystrophin after the 2OMeAO treatment. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (blue).

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

    Article Title: Systemic delivery of antisense oligoribonucleotide restores dystrophin expression in body-wide skeletal muscles

    doi: 10.1073/pnas.0406700102

    Figure Lengend Snippet: Restoration of dystrophin-associated proteins. ( Left ) Muscle of control mdx mouse. ( Right ) Muscle of mdx mouse after triple tail-vein injections of the 2OMeAO at weekly intervals. Membrane localized α-sarcoglycan, β-dystroglycan and neuronal nitric oxide synthase (nNOS) are detected in the fibers expressing dystrophin after the 2OMeAO treatment. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (blue).

    Article Snippet: Rabbit polyclonal antibodies to neuronal nitric oxide synthase and monoclonal antibodies to β-dystroglycan, α-sarcoglycan, and β-sarcoglycan were used according to the manufacturer's instructions (NovoCastra, Newcastle, U.K.).

    Techniques: Expressing

    Immunostaining for α-sarcoglycan and α-dystroglycan in cardiomyocytes. (a), Representative photomicrographs (×600, scale bar = 50μm) of immunostaining of α-sarcoglycan and α-dystroglycan in cardiomyocytes. (b), Quantitative analysis of immunostaining showed significantly increased staining of both α-sarcoglycan and α-dystroglycan in the HMGB1 group than the control group. HMGB1, high-mobility group box 1.

    Journal: PLoS ONE

    Article Title: The administration of high-mobility group box 1 fragment prevents deterioration of cardiac performance by enhancement of bone marrow mesenchymal stem cell homing in the delta-sarcoglycan-deficient hamster

    doi: 10.1371/journal.pone.0202838

    Figure Lengend Snippet: Immunostaining for α-sarcoglycan and α-dystroglycan in cardiomyocytes. (a), Representative photomicrographs (×600, scale bar = 50μm) of immunostaining of α-sarcoglycan and α-dystroglycan in cardiomyocytes. (b), Quantitative analysis of immunostaining showed significantly increased staining of both α-sarcoglycan and α-dystroglycan in the HMGB1 group than the control group. HMGB1, high-mobility group box 1.

    Article Snippet: The paraffin sections were used for immunohistochemistry and labeled using polyclonal CD31 antibody (1:50 CD31, Abcam, Cambridge, UK), anti-α-sarcoglycan (clone: Ad1/20A6; Novocastra, Weltzar, Germany), and anti-α-dystroglycan (clone: VIA4-1; Upstate Biotechnology, Lake Placid, NY) to assess capillary vascular density and the organization of cytoskeletal proteins.

    Techniques: Immunostaining, Staining

    Compound panel showing immunostaining of longitudinal sections of skeletal muscle fibres of patients with sensitive-motor polyneuropathy. The sections were immunolabelled with antibodies against α-sarcoglycan (A), β-sarcoglycan (B), γ-sarcoglycan

    Journal: Journal of Anatomy

    Article Title: Costameric proteins in human skeletal muscle during muscular inactivity

    doi: 10.1111/j.1469-7580.2008.00921.x

    Figure Lengend Snippet: Compound panel showing immunostaining of longitudinal sections of skeletal muscle fibres of patients with sensitive-motor polyneuropathy. The sections were immunolabelled with antibodies against α-sarcoglycan (A), β-sarcoglycan (B), γ-sarcoglycan

    Article Snippet: The following primary antibodies were used: anti-α-sarcoglycan diluted 1:100, anti-β-sarcoglycan diluted 1:200, anti-γ-sarcoglycan diluted 1:100, anti-δ-sarcoglycan diluted 1:50, and anti-dystrophin diluted 1:20 (all from Novocastra Laboratories, Newcastle Upon Tyne, Uk); anti-agrin diluted 1:100 (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA); anti-vinculin diluted 1:100, and anti-talin diluted 1:100 (both from Sigma Chemicals, St. Louis, MO, USA); anti-α7B-integrin diluted 1:50, anti-β1D-integrin diluted 1:50, and anti-α7A-integrin diluted 1:100 (synthetic peptides from the COOH terminal region; kindly provided by the laboratory of Professor Tarone, University of Turin).

    Techniques: Immunostaining

    Compound panel showing immunohistochemical findings in normal human skeletal muscle. Skeletal muscle fibres were immunolabelled with antibodies against α-sarcoglycan (A), β-sarcoglycan (B), γ-sarcoglycan (C), δ-sarcoglycan

    Journal: Journal of Anatomy

    Article Title: Costameric proteins in human skeletal muscle during muscular inactivity

    doi: 10.1111/j.1469-7580.2008.00921.x

    Figure Lengend Snippet: Compound panel showing immunohistochemical findings in normal human skeletal muscle. Skeletal muscle fibres were immunolabelled with antibodies against α-sarcoglycan (A), β-sarcoglycan (B), γ-sarcoglycan (C), δ-sarcoglycan

    Article Snippet: The following primary antibodies were used: anti-α-sarcoglycan diluted 1:100, anti-β-sarcoglycan diluted 1:200, anti-γ-sarcoglycan diluted 1:100, anti-δ-sarcoglycan diluted 1:50, and anti-dystrophin diluted 1:20 (all from Novocastra Laboratories, Newcastle Upon Tyne, Uk); anti-agrin diluted 1:100 (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA); anti-vinculin diluted 1:100, and anti-talin diluted 1:100 (both from Sigma Chemicals, St. Louis, MO, USA); anti-α7B-integrin diluted 1:50, anti-β1D-integrin diluted 1:50, and anti-α7A-integrin diluted 1:100 (synthetic peptides from the COOH terminal region; kindly provided by the laboratory of Professor Tarone, University of Turin).

    Techniques: Immunohistochemistry

    AAV-mediated microdystrophin expression restored DGC. A, Mock-treated and AAV-infected mdx hearts and age-matched BL10 heart were evaluated with monoclonal antibodies specific to N-terminal (human-specific) and C-terminal of dystrophin, β -sarcoglycan, and β -dystroglycan. Nuclei were counterstained with DAPI. Bar=100 μ m. B, Serial sections of virus-specific microdystrophin expression and recruitment of β -sarcoglycan and β -dystroglycan. DIC indicates differential interference contrast. Bar=200 μ m.

    Journal: Circulation

    Article Title: Microdystrophin Gene Therapy of Cardiomyopathy Restores Dystrophin-Glycoprotein Complex and Improves Sarcolemma Integrity in the Mdx Mouse Heart

    doi: 10.1161/01.CIR.0000089371.11664.27

    Figure Lengend Snippet: AAV-mediated microdystrophin expression restored DGC. A, Mock-treated and AAV-infected mdx hearts and age-matched BL10 heart were evaluated with monoclonal antibodies specific to N-terminal (human-specific) and C-terminal of dystrophin, β -sarcoglycan, and β -dystroglycan. Nuclei were counterstained with DAPI. Bar=100 μ m. B, Serial sections of virus-specific microdystrophin expression and recruitment of β -sarcoglycan and β -dystroglycan. DIC indicates differential interference contrast. Bar=200 μ m.

    Article Snippet: Additional monoclonal antibodies, including anti–dystrophin-C-terminal Dys-2 (1:30), anti– β -sarcoglycan (1:50), and anti– β -dystroglycan (1:50) antibodies (Novocastra) were used the same way as for Dys-3.

    Techniques: Expressing, Infection

    Functional and phenotypic correction in mdx mice following treatments with PNA AOs at 100 mg/kg for five weekly intravenous injections . ( a ) Serial immunostaining of dystrophin-associated protein complex (DAPC) in mdx mice. DAPC protein components β-dystroglycan, α-sarcoglycan, and β-sarcoglycan and neuronal nitric oxide synthase (nNOS) were detected in quadriceps from mdx mice treated with PNA AOs. Arrowheads point to the identical muscle fibers (bar = 200 μm). ( b ) Muscle function was assessed using grip strength test to determine the physical improvement of PNA AOs treated mdx mice. Significant force recovery was detected in treated mdx mice compared with untreated mdx controls ( t -test, * P

    Journal: Molecular Therapy. Nucleic Acids

    Article Title: Peptide Nucleic Acid Promotes Systemic Dystrophin Expression and Functional Rescue in Dystrophin-deficient mdx Mice

    doi: 10.1038/mtna.2015.27

    Figure Lengend Snippet: Functional and phenotypic correction in mdx mice following treatments with PNA AOs at 100 mg/kg for five weekly intravenous injections . ( a ) Serial immunostaining of dystrophin-associated protein complex (DAPC) in mdx mice. DAPC protein components β-dystroglycan, α-sarcoglycan, and β-sarcoglycan and neuronal nitric oxide synthase (nNOS) were detected in quadriceps from mdx mice treated with PNA AOs. Arrowheads point to the identical muscle fibers (bar = 200 μm). ( b ) Muscle function was assessed using grip strength test to determine the physical improvement of PNA AOs treated mdx mice. Significant force recovery was detected in treated mdx mice compared with untreated mdx controls ( t -test, * P

    Article Snippet: Rabbit polyclonal antibody to neuronal nitric oxide synthase and mouse monoclonal antibodies to β-dystroglycan, α-sarcoglycan, and β-sarcoglycan were used according to manufacturer's instructions (Novocastra, Newcastle upon Tyne, UK).

    Techniques: Functional Assay, Mouse Assay, Immunostaining

    HDAC4 directly represses structural gene expression via binding and inhibition of MEF2 activity. A ) Promoter sequence of γ-sarcoglycan spanning −812 to −861 nt upstream of transcriptional start. AT-rich MEF2 sites are indicated above, which were mutated singly in or combination for subsequent luciferase assays. B ) C2C12 myoblasts were tranfected with luciferase reporter constructs containing wild-type or MEF2 binding site deletion (ΔAT1, ΔAT2, or ΔAT1,2) promoters as indicated. Transfections occurred in the presence of MEF2 and/or HDAC4 as indicated. Luciferase activity was determined 48 h after transfection. C . D ) ChIP analysis was performed on C2C12 myotubes using either control IgG-, MEF2-, or HDAC4-specific antibodies. Bound DNA was analyzed by standard PCR generating a 300 bp fragment encompassing both MEF2 binding sites in the γ-sarcoglycan promoter. Input lane represents 5% of total starting material.

    Journal: The FASEB Journal

    Article Title: The deacetylase HDAC4 controls myocyte enhancing factor-2-dependent structural gene expression in response to neural activity

    doi: 10.1096/fj.08-115931

    Figure Lengend Snippet: HDAC4 directly represses structural gene expression via binding and inhibition of MEF2 activity. A ) Promoter sequence of γ-sarcoglycan spanning −812 to −861 nt upstream of transcriptional start. AT-rich MEF2 sites are indicated above, which were mutated singly in or combination for subsequent luciferase assays. B ) C2C12 myoblasts were tranfected with luciferase reporter constructs containing wild-type or MEF2 binding site deletion (ΔAT1, ΔAT2, or ΔAT1,2) promoters as indicated. Transfections occurred in the presence of MEF2 and/or HDAC4 as indicated. Luciferase activity was determined 48 h after transfection. C . D ) ChIP analysis was performed on C2C12 myotubes using either control IgG-, MEF2-, or HDAC4-specific antibodies. Bound DNA was analyzed by standard PCR generating a 300 bp fragment encompassing both MEF2 binding sites in the γ-sarcoglycan promoter. Input lane represents 5% of total starting material.

    Article Snippet: The following muscle specific antibodies were used: dystrophin (DYS1), β,γ-sarcoglycan, and desmin (Novocastra, Newcastle upon Tyne, UK); monoclonal myogenin antibody (Sigma, St. Louis, MO, USA); monoclonal caveolin-3 antibody (BD Biosciences, San Jose, CA, USA); and myotilin antibody (a kind gift from Mike Hauser, Duke University, Durham, NC, USA; ref. ).

    Techniques: Expressing, Binding Assay, Inhibition, Activity Assay, Sequencing, Luciferase, Construct, Transfection, Chromatin Immunoprecipitation, Polymerase Chain Reaction

    Generation of dystrophin/δ-sarcoglycan double mutant mice. ( A ) The breeding scheme. ( B ) Representative PCR genotyping results. Bkg, background; δSG KO, δ-sarcoglycan deficient mice; δ-Dko, δ-sarcoglycan/dystrophin

    Journal: Human Molecular Genetics

    Article Title: Sub-physiological sarcoglycan expression contributes to compensatory muscle protection in mdx mice

    doi: 10.1093/hmg/ddp015

    Figure Lengend Snippet: Generation of dystrophin/δ-sarcoglycan double mutant mice. ( A ) The breeding scheme. ( B ) Representative PCR genotyping results. Bkg, background; δSG KO, δ-sarcoglycan deficient mice; δ-Dko, δ-sarcoglycan/dystrophin

    Article Snippet: Dystrophin, utrophin, α-dystroglycan, α-Sarcoglycan, γ-Sarcoglycan and δ-Sarcoglycan were detected using the antibodies described earlier. β-Sarcoglycan was revealed with a mouse monoclonal antibody against the β-sarcoglycan N-terminus (NCL-b-SARC, 1:250; clone 5B1, IgG1; Novocastra).

    Techniques: Mutagenesis, Mouse Assay, Polymerase Chain Reaction

    Dystrophin/δ-sarcoglycan double knockout mice display a severer clinical phenotype. ( A ) Kyphosis (arrow) and hind limb joint contracture were apparent in δ-Dko mice. ( B ) Body weights of 3-month-old male BL10, mdx, δSG KO and δ-Dko

    Journal: Human Molecular Genetics

    Article Title: Sub-physiological sarcoglycan expression contributes to compensatory muscle protection in mdx mice

    doi: 10.1093/hmg/ddp015

    Figure Lengend Snippet: Dystrophin/δ-sarcoglycan double knockout mice display a severer clinical phenotype. ( A ) Kyphosis (arrow) and hind limb joint contracture were apparent in δ-Dko mice. ( B ) Body weights of 3-month-old male BL10, mdx, δSG KO and δ-Dko

    Article Snippet: Dystrophin, utrophin, α-dystroglycan, α-Sarcoglycan, γ-Sarcoglycan and δ-Sarcoglycan were detected using the antibodies described earlier. β-Sarcoglycan was revealed with a mouse monoclonal antibody against the β-sarcoglycan N-terminus (NCL-b-SARC, 1:250; clone 5B1, IgG1; Novocastra).

    Techniques: Double Knockout, Mouse Assay

    The muscle biopsy of the patient with the genetically confirmed LGMD2D showing no sign of dystrophy (A) and normal expression of all sarcoglycans (B, C, D, E) . The biopsy was taken at the age of 8 years.

    Journal: Acta Myologica

    Article Title: Childhood onset limb-girdle muscular dystrophies in the Aegean part of Turkey

    doi:

    Figure Lengend Snippet: The muscle biopsy of the patient with the genetically confirmed LGMD2D showing no sign of dystrophy (A) and normal expression of all sarcoglycans (B, C, D, E) . The biopsy was taken at the age of 8 years.

    Article Snippet: Spectrin (Novo-castra, UK, NCL-spec1), dystrophin N-terminus (Novo-castra, UK, NCL-dys3), adhalin (Novocastra, UK, NCL-a-sarc), other sarcoglycans (beta, delta, gamma; Novo-castra, UK, NCL-b-d-g-sarc), laminin alpha-2 chain (Novo-castra, UK, NCL-merosin), myotilin (Novo-castra, UK, NCL-myotilin), collagen VI (Novocastra, UK, NCL-COLL-vI), β-dystroglycan (Novocastra, UK, NCL-b-DG), HLA Class 1 (Novo-castra, UK, NCL-HLA-ABC), NCAM (ThermoScientific, CA, USA, CD56), nitric oxide synthase-1 (Novo-castra, UK, NCLNOS-1), emerin (Novo-castra, UK, NCL-emerin), caveolin 3 (Novus Biologicals, CA, USA, NB110-5029), calpain 3 (Abcam, Cambridge, UK, ab103250) and dysferlin (Novo-castra, UK, NCL-Hamlet-2) antibodies were used by standard techniques for immunohistochemical analyses.

    Techniques: Expressing

    Disruption of dystrophin expression in the skeletal muscle of DMD KO rabbits. (A-C) Immunofluorescence staining of muscle sections from WT and DMD KO rabbits with mouse monoclonal antibodies against dystrophin (A), glycosylated α-dystroglycan (B) and α-sarcoglycan (C). Nuclei were stained by DAPI. Scale bar: 100 µm.

    Journal: Disease Models & Mechanisms

    Article Title: A novel rabbit model of Duchenne muscular dystrophy generated by CRISPR/Cas9

    doi: 10.1242/dmm.032201

    Figure Lengend Snippet: Disruption of dystrophin expression in the skeletal muscle of DMD KO rabbits. (A-C) Immunofluorescence staining of muscle sections from WT and DMD KO rabbits with mouse monoclonal antibodies against dystrophin (A), glycosylated α-dystroglycan (B) and α-sarcoglycan (C). Nuclei were stained by DAPI. Scale bar: 100 µm.

    Article Snippet: Primary antibodies against dystrophin (MANDYS1 clone 3B7, 1:20, Developmental Studies Hybridoma Bank), α-dystroglycan (sc-53987, 1:50, Santa Cruz Biotechnology), α-sarcoglycan (NCL-L-a-SARC, 1:100, Leica Biosystems) and caveolin 3 (610420, 1:500, BD Biosciences) were used.

    Techniques: Expressing, Immunofluorescence, Staining

    Tsp expression in muscle of Drosophila rescues MD due to deletion of the δ-sarcoglycan-like gene (Sgcd 840 ). ( A ) Fly survival was compared over a period of 40 days. p

    Journal: eLife

    Article Title: Thrombospondin expression in myofibers stabilizes muscle membranes

    doi: 10.7554/eLife.17589

    Figure Lengend Snippet: Tsp expression in muscle of Drosophila rescues MD due to deletion of the δ-sarcoglycan-like gene (Sgcd 840 ). ( A ) Fly survival was compared over a period of 40 days. p

    Article Snippet: Primary antibodies included: δ-sarcoglycan (Abcam, ab92896, 1:100), α-sarcoglycan, β-sarcoglycan, γ-sarcoglycan (NovaCastra, Buffalo Grove, IL, NCL-a-sarc, NCL-b-sarc and NCL-g-sarc, all 1:250), β-dystroglycan (Development Studies Hybridoma Bank, Iowa City IA, MANDAG2; 1:50), utrophin (Santa Cruz Biotechnology, MANCHO7, sc-81557; 1:50), dystrophin (Abcam, ab15277, 1:200) and β1D-integrin (Millipore, Billerica, MA, MAB1900, 1:250).

    Techniques: Expressing

    Thbs4 enhances stabilizing proteins at the sarcolemma and directly interacts with integrins. ( A , B ) Representative Western blots of sarcolemmal protein extracts from the quadriceps of the indicated groups of mice at three months of age (n = 4–5 biological replicates). Sgcd -/- Tg and mdx Tg indicate Sgcd -/- and mdx with skeletal muscle specific Thbs4 overexpression, respectively. Ponceau staining of a nonspecific band and dihydropyridine receptor α1 (Cav1.1) were used as loading controls. Abbreviations: Utro, utrophin; Dystro, dystrophin; Dysfer, dysferlin; α-DG, α-dystroglycan; β-DG, β-dystroglycan; δ-SCG, δ-sarcoglycan; α-SCG, α-sarcoglycan; β-SGC, β-sarcoglycan; β1D-, α7- and α5-itg (integrin). ( C ) Immunoblots for β1D- and α7-Integrin (Itg), β-dystroglycan (DG) and Thbs4 (Flag) from neonatal rat ventricular myocyte extracts immunoprecipitated with a Flag antibody (Thbs4). Adβgal was used as a control infection (n = 3 biological replicates). An adenovirus expressing a Flag-tagged Thbs4 protein was used to achieve high level of this protein to identify the interaction. ( D , E ) Representative Western blots for Thbs4, α5- and β1D-integrin (itg) from intracellular vesicular isolates from WT and Thbs4 Tg quadriceps ( D ) or WT and Sgcd -/- quadriceps ( E ) that were immunoprecipitated with an antibody raised against the cytoplasmic domain of β1D-integrin (n = 3 biological replicates), showing that Thbs4 and α5-integrin localize to β1D-integrin-positive intracellular vesicles. α-tubulin and Gapdh are presented as loading control. DOI: http://dx.doi.org/10.7554/eLife.17589.019

    Journal: eLife

    Article Title: Thrombospondin expression in myofibers stabilizes muscle membranes

    doi: 10.7554/eLife.17589

    Figure Lengend Snippet: Thbs4 enhances stabilizing proteins at the sarcolemma and directly interacts with integrins. ( A , B ) Representative Western blots of sarcolemmal protein extracts from the quadriceps of the indicated groups of mice at three months of age (n = 4–5 biological replicates). Sgcd -/- Tg and mdx Tg indicate Sgcd -/- and mdx with skeletal muscle specific Thbs4 overexpression, respectively. Ponceau staining of a nonspecific band and dihydropyridine receptor α1 (Cav1.1) were used as loading controls. Abbreviations: Utro, utrophin; Dystro, dystrophin; Dysfer, dysferlin; α-DG, α-dystroglycan; β-DG, β-dystroglycan; δ-SCG, δ-sarcoglycan; α-SCG, α-sarcoglycan; β-SGC, β-sarcoglycan; β1D-, α7- and α5-itg (integrin). ( C ) Immunoblots for β1D- and α7-Integrin (Itg), β-dystroglycan (DG) and Thbs4 (Flag) from neonatal rat ventricular myocyte extracts immunoprecipitated with a Flag antibody (Thbs4). Adβgal was used as a control infection (n = 3 biological replicates). An adenovirus expressing a Flag-tagged Thbs4 protein was used to achieve high level of this protein to identify the interaction. ( D , E ) Representative Western blots for Thbs4, α5- and β1D-integrin (itg) from intracellular vesicular isolates from WT and Thbs4 Tg quadriceps ( D ) or WT and Sgcd -/- quadriceps ( E ) that were immunoprecipitated with an antibody raised against the cytoplasmic domain of β1D-integrin (n = 3 biological replicates), showing that Thbs4 and α5-integrin localize to β1D-integrin-positive intracellular vesicles. α-tubulin and Gapdh are presented as loading control. DOI: http://dx.doi.org/10.7554/eLife.17589.019

    Article Snippet: Primary antibodies included: δ-sarcoglycan (Abcam, ab92896, 1:100), α-sarcoglycan, β-sarcoglycan, γ-sarcoglycan (NovaCastra, Buffalo Grove, IL, NCL-a-sarc, NCL-b-sarc and NCL-g-sarc, all 1:250), β-dystroglycan (Development Studies Hybridoma Bank, Iowa City IA, MANDAG2; 1:50), utrophin (Santa Cruz Biotechnology, MANCHO7, sc-81557; 1:50), dystrophin (Abcam, ab15277, 1:200) and β1D-integrin (Millipore, Billerica, MA, MAB1900, 1:250).

    Techniques: Western Blot, Mouse Assay, Over Expression, Staining, Immunoprecipitation, Infection, Expressing

    Generation of dystrophin/δ-sarcoglycan double mutant mice. ( A ) The breeding scheme. ( B ) Representative PCR genotyping results. Bkg, background; δSG KO, δ-sarcoglycan deficient mice; δ-Dko, δ-sarcoglycan/dystrophin

    Journal: Human Molecular Genetics

    Article Title: Sub-physiological sarcoglycan expression contributes to compensatory muscle protection in mdx mice

    doi: 10.1093/hmg/ddp015

    Figure Lengend Snippet: Generation of dystrophin/δ-sarcoglycan double mutant mice. ( A ) The breeding scheme. ( B ) Representative PCR genotyping results. Bkg, background; δSG KO, δ-sarcoglycan deficient mice; δ-Dko, δ-sarcoglycan/dystrophin

    Article Snippet: Utrophin was examined with a mouse monoclonal antibody against the utrophin N-terminal domain (VP-U579, 1:20; clone DRP3/20C5, IgG1; Vector Laboratories, Burlingame, CA). β-Dystroglycan was revealed with NCL-b-DG (1:50; Novocastra). α-Sarcoglycan was detected with VP-A105 (1:50; Vector Laboratories). β-Sarcoglycan was revealed with NCL-b-SARC (1:50; Novocastra). γ-Sarcoglycan was detected with VP-G803 (1:50; Vector Laboratories). δ-Sarcoglycan was detected with sc28281 (1:50; Santa Cruz Biotechnology Inc.).

    Techniques: Mutagenesis, Mouse Assay, Polymerase Chain Reaction

    Dystrophin/δ-sarcoglycan double knockout mice display a severer clinical phenotype. ( A ) Kyphosis (arrow) and hind limb joint contracture were apparent in δ-Dko mice. ( B ) Body weights of 3-month-old male BL10, mdx, δSG KO and δ-Dko

    Journal: Human Molecular Genetics

    Article Title: Sub-physiological sarcoglycan expression contributes to compensatory muscle protection in mdx mice

    doi: 10.1093/hmg/ddp015

    Figure Lengend Snippet: Dystrophin/δ-sarcoglycan double knockout mice display a severer clinical phenotype. ( A ) Kyphosis (arrow) and hind limb joint contracture were apparent in δ-Dko mice. ( B ) Body weights of 3-month-old male BL10, mdx, δSG KO and δ-Dko

    Article Snippet: Utrophin was examined with a mouse monoclonal antibody against the utrophin N-terminal domain (VP-U579, 1:20; clone DRP3/20C5, IgG1; Vector Laboratories, Burlingame, CA). β-Dystroglycan was revealed with NCL-b-DG (1:50; Novocastra). α-Sarcoglycan was detected with VP-A105 (1:50; Vector Laboratories). β-Sarcoglycan was revealed with NCL-b-SARC (1:50; Novocastra). γ-Sarcoglycan was detected with VP-G803 (1:50; Vector Laboratories). δ-Sarcoglycan was detected with sc28281 (1:50; Santa Cruz Biotechnology Inc.).

    Techniques: Double Knockout, Mouse Assay

    Overexpression of Serpina3n restores the abundance of sarcolemma adhesion components in two dystrophic mouse models. ( A ) Immunohistochemistry of transverse quadriceps histological sections or ( B ) western blotting from quadriceps muscle of membrane protein extracts from mice at 2–3 months of age. The genotypes of mice used are shown with the indicated antibodies. Nuclei are stained in blue with DAPI, and Ponceau was used as a western loading control. Dystr. dystrophin; Utr, utrophin; β1D-Itg, β1D-integrin; α7-Itg, α7-integrin; δ-SG, δ-sarcoglycan; α-DG, α-dystroglycan; β-DG, β-dystroglycan; β-SG, β-sarcoglycan; α-SG; α-sarcoglycan; CaV1.1, L-type calcium channel. ( C ) Quantitative protein analysis for the indicated proteins in dystrophic muscle following overexpression of Serpina3n. A total of three separate animals for each genotype were used for densitometric analysis. * P

    Journal: Human Molecular Genetics

    Article Title: Genetic overexpression of Serpina3n attenuates muscular dystrophy in mice

    doi: 10.1093/hmg/ddw005

    Figure Lengend Snippet: Overexpression of Serpina3n restores the abundance of sarcolemma adhesion components in two dystrophic mouse models. ( A ) Immunohistochemistry of transverse quadriceps histological sections or ( B ) western blotting from quadriceps muscle of membrane protein extracts from mice at 2–3 months of age. The genotypes of mice used are shown with the indicated antibodies. Nuclei are stained in blue with DAPI, and Ponceau was used as a western loading control. Dystr. dystrophin; Utr, utrophin; β1D-Itg, β1D-integrin; α7-Itg, α7-integrin; δ-SG, δ-sarcoglycan; α-DG, α-dystroglycan; β-DG, β-dystroglycan; β-SG, β-sarcoglycan; α-SG; α-sarcoglycan; CaV1.1, L-type calcium channel. ( C ) Quantitative protein analysis for the indicated proteins in dystrophic muscle following overexpression of Serpina3n. A total of three separate animals for each genotype were used for densitometric analysis. * P

    Article Snippet: Primary antibodies used included: dystrophin [MANDYS1(3B7) 1:500]; utrophin (DSHB Mancho3-8A4 1:500); α7-integrin (Santa Cruz SC-27706 1:500); β1D-integrin (Chemicon; MAB1900, 1:200); δ-sarcoglycan (Abcam ab137101 1:1000); α-dystroglycan (Millipore 05-593 1:500); β-dystroglycan (MANDAG2 clone 7D11 1:500); β-sarcoglycan (Vector labs VP-B206 1: 500); α-sarcoglycan (DSHB IVD3(1)A9 1:500) and CaV1.1 (Pierce MA3-920 1:1000).

    Techniques: Over Expression, Immunohistochemistry, Western Blot, Mouse Assay, Staining

    Differences in α-dystrobrevin splice variants between cardiac and skeletal muscle DAPC. ( A ) Western blot analysis of α-dystrobrevins in mouse cardiac (C) and skeletal (S) muscle total protein lysates, DYS-IPs and IgG-IPs. α3-dystrobrevin associates with dystrophin in the heart. Fold differences in α-dystrobrevin abundance in cardiac vs. skeletal muscle DYS-IPs relative to dystrophin are shown (averages ±SD, N = 3). ( B ) Immunolabeling of wild type cardiac sections for α1 and α2-dystrobrevins. Scale bar: 50 µm. ( C ) Immunolabeling of mdx cardiac tissue section for β-sarcoglycan. Scale bar: 50 µm. ( D ) Western blot analysis of α-dystrobrevins in heart protein lysates from wild type (WT) and mdx mice. Fold differences in α-dystrobrevin abundance in WT vs. mdx cardiac lysates relative to GAPDH are shown (averages ±SD, N = 3) ( E ) Western blot analysis of α-dystrobrevins in DYS-IPs from human (H) and mouse (M) cardiac samples. Additional α-dystrobrevin isoforms (arrow heads) are detected in human cardiac lysates and DYS-IP.

    Journal: PLoS ONE

    Article Title: Proteomic Analysis Reveals New Cardiac-Specific Dystrophin-Associated Proteins

    doi: 10.1371/journal.pone.0043515

    Figure Lengend Snippet: Differences in α-dystrobrevin splice variants between cardiac and skeletal muscle DAPC. ( A ) Western blot analysis of α-dystrobrevins in mouse cardiac (C) and skeletal (S) muscle total protein lysates, DYS-IPs and IgG-IPs. α3-dystrobrevin associates with dystrophin in the heart. Fold differences in α-dystrobrevin abundance in cardiac vs. skeletal muscle DYS-IPs relative to dystrophin are shown (averages ±SD, N = 3). ( B ) Immunolabeling of wild type cardiac sections for α1 and α2-dystrobrevins. Scale bar: 50 µm. ( C ) Immunolabeling of mdx cardiac tissue section for β-sarcoglycan. Scale bar: 50 µm. ( D ) Western blot analysis of α-dystrobrevins in heart protein lysates from wild type (WT) and mdx mice. Fold differences in α-dystrobrevin abundance in WT vs. mdx cardiac lysates relative to GAPDH are shown (averages ±SD, N = 3) ( E ) Western blot analysis of α-dystrobrevins in DYS-IPs from human (H) and mouse (M) cardiac samples. Additional α-dystrobrevin isoforms (arrow heads) are detected in human cardiac lysates and DYS-IP.

    Article Snippet: Antibodies to DAPC members are: isoform specific anti- α1-, β1- or β2-syntrophin, and anti- α1- or α2-dystrobrevin antibodies , , ; pan anti-syntrophin (ab11425, Abcam); Manex1011B to dystrophin and MANDAG2 to β-dystroglycan (DSHB); clone IIH6C4 to α-dystroglycan (Upstate); anti-nNOS (#610308) and anti-α-dystrobrevin (#610766, BD Bioscience); anti-β-sarcoglycan (clone 5B1, Leica Microsystems).

    Techniques: Western Blot, Immunolabeling, Mouse Assay

    Dependence of ecto-nucleotidase activity of α-sarcoglycan-expressing HEK-293 cells on the bivalent cation concentrations ( A ) ATP-hydrolysing activity was measured in stably transfected cells in the presence of 4 mM Mg 2+ or 2 mM Ca 2+ or both (4 mM Mg 2+ and 2 mM Ca 2+ ). ( B ) ATP-hydrolysing activity of α-sarcoglycan stably transfected HEK-293 cells was measured either in the presence of 4 mM Mg 2+ and the indicated concentrations of Ca 2+ (○) or in the presence of 2 mM Ca 2+ and the indicated concentrations of Mg 2+ (•). The plot shows the values after the subtraction of the empty vector-transfected activity. Results are from four experiments performed in triplicate. ** P

    Journal: Biochemical Journal

    Article Title: Characterization of the ATP-hydrolysing activity of ?-sarcoglycan

    doi: 10.1042/BJ20031644

    Figure Lengend Snippet: Dependence of ecto-nucleotidase activity of α-sarcoglycan-expressing HEK-293 cells on the bivalent cation concentrations ( A ) ATP-hydrolysing activity was measured in stably transfected cells in the presence of 4 mM Mg 2+ or 2 mM Ca 2+ or both (4 mM Mg 2+ and 2 mM Ca 2+ ). ( B ) ATP-hydrolysing activity of α-sarcoglycan stably transfected HEK-293 cells was measured either in the presence of 4 mM Mg 2+ and the indicated concentrations of Ca 2+ (○) or in the presence of 2 mM Ca 2+ and the indicated concentrations of Mg 2+ (•). The plot shows the values after the subtraction of the empty vector-transfected activity. Results are from four experiments performed in triplicate. ** P

    Article Snippet: The proteins (40–50 μg/lane) were resolved by SDS/PAGE, blotted on to a nitrocellulose membrane and examined with the following antibodies: mouse monoclonal anti-α-sarcoglycan (NCL-a-SARC, 1:500), anti-dystrophin (NCL-DYS2, 1:100), and anti-β-dystroglycan (NCL-b-DG, 1:100) from Novocastra and rabbit polyclonal anti-β-actin (AC-15, 1:4000) from Sigma.

    Techniques: Activity Assay, Expressing, Stable Transfection, Transfection, Plasmid Preparation

    Transient transfection of HEK-293 cells with α-sarcoglycan ( A ) Western-blot analysis of rabbit skeletal-muscle sarcolemma (rabbit SL), and HEK-293 cell lysates from α-sarcoglycan-transiently transfected cells (α-SG), empty vector (pcDNA3) transfected cells and untransfected cells (wild-type) by using an antibody specific to α-sarcoglycan. ( B ) Western-blot analysis of rabbit skeletal-muscle sarcolemma (rabbit SL), and HEK-293 cell lysates from α-sarcoglycan-transiently transfected cells (HEK α-SG) either untreated (left two lanes) or treated with N -glycosidase F (right two lanes). Amounts of protein from HEK-293 cell lysates were estimated ( A , B ) by using an antibody against β-actin. ( C ) Phase-contrast image (left panel) and immunofluorescence staining of non-permeabilized α-sarcoglycan transiently transfected HEK-293 cells with the antibody directed against the extracellular domain of α-sarcoglycan (right panel). ( D ) Extracellular ATP hydrolysis of wild-type HEK-293 cells (wt), or cells transiently transfected with the empty vector (pcDNA3) or with α-sarcoglycan (α-SG). Results are from three experiments performed in triplicate; ** P

    Journal: Biochemical Journal

    Article Title: Characterization of the ATP-hydrolysing activity of ?-sarcoglycan

    doi: 10.1042/BJ20031644

    Figure Lengend Snippet: Transient transfection of HEK-293 cells with α-sarcoglycan ( A ) Western-blot analysis of rabbit skeletal-muscle sarcolemma (rabbit SL), and HEK-293 cell lysates from α-sarcoglycan-transiently transfected cells (α-SG), empty vector (pcDNA3) transfected cells and untransfected cells (wild-type) by using an antibody specific to α-sarcoglycan. ( B ) Western-blot analysis of rabbit skeletal-muscle sarcolemma (rabbit SL), and HEK-293 cell lysates from α-sarcoglycan-transiently transfected cells (HEK α-SG) either untreated (left two lanes) or treated with N -glycosidase F (right two lanes). Amounts of protein from HEK-293 cell lysates were estimated ( A , B ) by using an antibody against β-actin. ( C ) Phase-contrast image (left panel) and immunofluorescence staining of non-permeabilized α-sarcoglycan transiently transfected HEK-293 cells with the antibody directed against the extracellular domain of α-sarcoglycan (right panel). ( D ) Extracellular ATP hydrolysis of wild-type HEK-293 cells (wt), or cells transiently transfected with the empty vector (pcDNA3) or with α-sarcoglycan (α-SG). Results are from three experiments performed in triplicate; ** P

    Article Snippet: The proteins (40–50 μg/lane) were resolved by SDS/PAGE, blotted on to a nitrocellulose membrane and examined with the following antibodies: mouse monoclonal anti-α-sarcoglycan (NCL-a-SARC, 1:500), anti-dystrophin (NCL-DYS2, 1:100), and anti-β-dystroglycan (NCL-b-DG, 1:100) from Novocastra and rabbit polyclonal anti-β-actin (AC-15, 1:4000) from Sigma.

    Techniques: Transfection, Western Blot, Plasmid Preparation, Immunofluorescence, Staining

    Ecto-nucleotidase activity of HEK-293 cells stably transfected with α-sarcoglycan ( A ) ATP-hydrolysing activity, measured as indicated in the Experimental section, was performed in HEK-293 cells transfected with the empty vector (pcDNA3) or with α-sarcoglycan (α-SG). Results are from four experiments performed in triplicate; ** P

    Journal: Biochemical Journal

    Article Title: Characterization of the ATP-hydrolysing activity of ?-sarcoglycan

    doi: 10.1042/BJ20031644

    Figure Lengend Snippet: Ecto-nucleotidase activity of HEK-293 cells stably transfected with α-sarcoglycan ( A ) ATP-hydrolysing activity, measured as indicated in the Experimental section, was performed in HEK-293 cells transfected with the empty vector (pcDNA3) or with α-sarcoglycan (α-SG). Results are from four experiments performed in triplicate; ** P

    Article Snippet: The proteins (40–50 μg/lane) were resolved by SDS/PAGE, blotted on to a nitrocellulose membrane and examined with the following antibodies: mouse monoclonal anti-α-sarcoglycan (NCL-a-SARC, 1:500), anti-dystrophin (NCL-DYS2, 1:100), and anti-β-dystroglycan (NCL-b-DG, 1:100) from Novocastra and rabbit polyclonal anti-β-actin (AC-15, 1:4000) from Sigma.

    Techniques: Activity Assay, Stable Transfection, Transfection, Plasmid Preparation

    Effects of antibodies against α-sarcoglycan on the ecto-nucleotidase activity of HEK-293 cells stably expressing the protein ATP-hydrolysing activity of cells incubated in the absence or in the presence of a monoclonal antibody against the extracellular domain of α-sarcoglycan encompassing the ATP-binding site (+mAb) or a polyclonal antibody specific for the C-terminal domain of the protein (+pAb). HEK-293 cells were preincubated for 30 min at 4 °C with or without antibodies. Results are from four experiments performed in triplicate; ** P

    Journal: Biochemical Journal

    Article Title: Characterization of the ATP-hydrolysing activity of ?-sarcoglycan

    doi: 10.1042/BJ20031644

    Figure Lengend Snippet: Effects of antibodies against α-sarcoglycan on the ecto-nucleotidase activity of HEK-293 cells stably expressing the protein ATP-hydrolysing activity of cells incubated in the absence or in the presence of a monoclonal antibody against the extracellular domain of α-sarcoglycan encompassing the ATP-binding site (+mAb) or a polyclonal antibody specific for the C-terminal domain of the protein (+pAb). HEK-293 cells were preincubated for 30 min at 4 °C with or without antibodies. Results are from four experiments performed in triplicate; ** P

    Article Snippet: The proteins (40–50 μg/lane) were resolved by SDS/PAGE, blotted on to a nitrocellulose membrane and examined with the following antibodies: mouse monoclonal anti-α-sarcoglycan (NCL-a-SARC, 1:500), anti-dystrophin (NCL-DYS2, 1:100), and anti-β-dystroglycan (NCL-b-DG, 1:100) from Novocastra and rabbit polyclonal anti-β-actin (AC-15, 1:4000) from Sigma.

    Techniques: Activity Assay, Stable Transfection, Expressing, Incubation, Binding Assay

    Western-blot analysis and total ecto-nucleotidase activity of C2C12 cells ( A ) Western-blot analysis of cell lysates obtained from proliferating (P), confluent (C) and myotubes at different days of culture. Lysate proteins (50 μg) were separated on 10% SDS/polyacrylamide slab gels, blotted on nitrocellulose filters and probed with specific antibodies, as indicated. ( B ) ATP-hydrolysing activity of C2C12 myoblasts (Mb) and 4- and 8-day-old myotubes (4d-Mt and 8d-Mt respectively) was determined by measuring P i ]. Cells were incubated for 15 min in a medium with the following composition: 20 mM Hepes, 140 mM NaCl, 4 mM MgCl 2 and 2 mM CaCl 2 . The reaction was started by adding 4 mM ATP and terminated by withdrawing a supernatant aliquot. Results are from four experiments performed in triplicate. ( C ) Effect of polyclonal (+pAb) and monoclonal (+mAb) antibodies specific for α-sarcoglycan on total ecto-ATPase activity of 7-day-old C2C12 myotubes incubated 30 min as above. Results are from three experiments performed in triplicate. * P

    Journal: Biochemical Journal

    Article Title: Characterization of the ATP-hydrolysing activity of ?-sarcoglycan

    doi: 10.1042/BJ20031644

    Figure Lengend Snippet: Western-blot analysis and total ecto-nucleotidase activity of C2C12 cells ( A ) Western-blot analysis of cell lysates obtained from proliferating (P), confluent (C) and myotubes at different days of culture. Lysate proteins (50 μg) were separated on 10% SDS/polyacrylamide slab gels, blotted on nitrocellulose filters and probed with specific antibodies, as indicated. ( B ) ATP-hydrolysing activity of C2C12 myoblasts (Mb) and 4- and 8-day-old myotubes (4d-Mt and 8d-Mt respectively) was determined by measuring P i ]. Cells were incubated for 15 min in a medium with the following composition: 20 mM Hepes, 140 mM NaCl, 4 mM MgCl 2 and 2 mM CaCl 2 . The reaction was started by adding 4 mM ATP and terminated by withdrawing a supernatant aliquot. Results are from four experiments performed in triplicate. ( C ) Effect of polyclonal (+pAb) and monoclonal (+mAb) antibodies specific for α-sarcoglycan on total ecto-ATPase activity of 7-day-old C2C12 myotubes incubated 30 min as above. Results are from three experiments performed in triplicate. * P

    Article Snippet: The proteins (40–50 μg/lane) were resolved by SDS/PAGE, blotted on to a nitrocellulose membrane and examined with the following antibodies: mouse monoclonal anti-α-sarcoglycan (NCL-a-SARC, 1:500), anti-dystrophin (NCL-DYS2, 1:100), and anti-β-dystroglycan (NCL-b-DG, 1:100) from Novocastra and rabbit polyclonal anti-β-actin (AC-15, 1:4000) from Sigma.

    Techniques: Western Blot, Activity Assay, Incubation

    The expression analyses of α-, β-, γ-sarcoglycan and dysferlin protein in the muscles of patient by Western blot. The total proteins extracted from skeletal muscles of normal person (lane 1), relative normal muscle biopsies of patient IV-7 (lane 2), and dystrophic muscle biopsies of patient IV-7 (lane 3).

    Journal: Journal of Translational Medicine

    Article Title: Differential expression profiling between the relative normal and dystrophic muscle tissues from the same LGMD patient

    doi: 10.1186/1479-5876-4-53

    Figure Lengend Snippet: The expression analyses of α-, β-, γ-sarcoglycan and dysferlin protein in the muscles of patient by Western blot. The total proteins extracted from skeletal muscles of normal person (lane 1), relative normal muscle biopsies of patient IV-7 (lane 2), and dystrophic muscle biopsies of patient IV-7 (lane 3).

    Article Snippet: Antibodies and Western blot Antibodies against Dysferlin (E20), α-Sarcoglycan (D-20), β-Sarcoglycan (A-17), γ-Sarcoglycan (D-18), α-Tubulin, and horseradish peroxidase (HRP)-conjugated second antibodies were from Santa Cruz Biotechnology.

    Techniques: Expressing, Western Blot