sarcoglycan complex  (Novocastra)

Journal: Molecular Therapy. Nucleic Acids

Article Title: Long-Term Efficacy of Systemic Multiexon Skipping Targeting Dystrophin Exons 45–55 With a Cocktail of Vivo-Morpholinos in Mdx52 Mice

doi: 10.1038/mtna.2014.76

Figure Lengend Snippet: I mmunohistochemistry of dystrophin-associated proteins accompanied after 9 times i.v. injections of the 10-vPMO cocktail. Dystrophin, α1-syntrophin, nNOS, α-sarcoglycan, and β-dystroglycan were stained on serial sections of the muscles form nontreated and treated mdx52 mice. Biceps femoris muscles were shown as representative data in 3 treated mice. Asterisks indicate the same muscle fiber between the images. Scale bar, 100 μm.

Article Snippet: In systemic treatment with the 10-vPMO cocktail, the diaphragm, biceps femoris, quadriceps, gastrocnemius, tibialis anterior, biceps brachii, triceps brachii, and heart muscles were examined 2 weeks after the final injection using anti-dystrophin (P7) antibody and antibodies against dystrophin-associated proteins: anti-α1-syntrophin rabbit polyclonal antibody (1:200, Abcam, Cambridge, UK), anti-nNOS rabbit polyclonal antibody (1:100, Invitrogen), anti-α-sarcoglycan mouse monoclonal antibody (1:10, Novocastra Laboratories), and anti-β-dystroglycan mouse monoclonal antibody (1:5, Novocastra Laboratories).

Techniques: Staining, Mouse Assay

Journal: Cardiovascular Diabetology

Article Title: Successful metabolic adaptations leading to the prevention of high fat diet-induced murine cardiac remodeling

doi: 10.1186/s12933-015-0286-0

Figure Lengend Snippet: Hexokinase II is also reduced in the MRL cardiac tissue. a Representative immunoblot of HKII in the heart tissues. b γ-Sarcoglycan loading control. c Quantification of HKII normalized to γ-sarcoglycan and B6 CD quantities (N = 12, B6 CD verses MRL CD p = 0.000, B6 HFD versus MRL HFD p = 0.001, and B6 CD versus B6 HFD p = 0.013)

Article Snippet: The membranes were incubated at 4 °C overnight in 5 % NFDM/TBST with primary antibodies [anti-AMPK, anti-pAMPK, and PGC-1α (1:1000, Cell Signaling Technology, Danvers, MA, USA); anti-ACC, anti-pACC, fibronectin (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-HKII (1:5000, Millipore, Billerica, MA, USA); VDAC, Oxidative Phosphorylation antibody cocktail (1:1000, Abcam, Cambridge, MA, USA), Glut4 (1:500, EMD Millipore, Darmstadt, Germany), and γ-sarcoglycan (1:1000, Novocastra Labs, Buffalo Grove, IL, USA)].

Techniques:

Journal: Cardiovascular Diabetology

Article Title: Successful metabolic adaptations leading to the prevention of high fat diet-induced murine cardiac remodeling

doi: 10.1186/s12933-015-0286-0

Figure Lengend Snippet: Mitochondrial differences in the two mouse strains. a The hearts from MRL mice contain increased mitochondrial genomes when normalized to nuclear DNA (N = 14, CD comparisons p = 0.002 and HFD comparisons p = 0.043). b However, quantification of VDAC immunoblots demonstrated no change in the outer mitochondrial membrane protein (N = 6). c Control to γ-sarcoglycan immunoblot. d VDAC normalized to γ-sarcoglycan (N = 6)

Article Snippet: The membranes were incubated at 4 °C overnight in 5 % NFDM/TBST with primary antibodies [anti-AMPK, anti-pAMPK, and PGC-1α (1:1000, Cell Signaling Technology, Danvers, MA, USA); anti-ACC, anti-pACC, fibronectin (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-HKII (1:5000, Millipore, Billerica, MA, USA); VDAC, Oxidative Phosphorylation antibody cocktail (1:1000, Abcam, Cambridge, MA, USA), Glut4 (1:500, EMD Millipore, Darmstadt, Germany), and γ-sarcoglycan (1:1000, Novocastra Labs, Buffalo Grove, IL, USA)].

Techniques: Mouse Assay, Western Blot

Journal: Cardiovascular Diabetology

Article Title: Successful metabolic adaptations leading to the prevention of high fat diet-induced murine cardiac remodeling

doi: 10.1186/s12933-015-0286-0

Figure Lengend Snippet: Decreased MRL cardiac ACC indicates increased fatty acid metabolism. a pACC immunoblot. b Quantification normalized to γ-sarcoglycan shows the MRL tissues have reduced pACC (N = 6, B6 HFD versus MRL HFD p = 0.011). c ACC immunoblot. d Quantification normalized to γ-sarcoglycan shows the MRL tissues have reduced ACC (N = 6, B6 HFD versus MRL HFD p = 0.033, B6 CD versus B6 HFD p = 0.013)

Article Snippet: The membranes were incubated at 4 °C overnight in 5 % NFDM/TBST with primary antibodies [anti-AMPK, anti-pAMPK, and PGC-1α (1:1000, Cell Signaling Technology, Danvers, MA, USA); anti-ACC, anti-pACC, fibronectin (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-HKII (1:5000, Millipore, Billerica, MA, USA); VDAC, Oxidative Phosphorylation antibody cocktail (1:1000, Abcam, Cambridge, MA, USA), Glut4 (1:500, EMD Millipore, Darmstadt, Germany), and γ-sarcoglycan (1:1000, Novocastra Labs, Buffalo Grove, IL, USA)].

Techniques:

Journal: Cardiovascular Diabetology

Article Title: Successful metabolic adaptations leading to the prevention of high fat diet-induced murine cardiac remodeling

doi: 10.1186/s12933-015-0286-0

Figure Lengend Snippet: The hearts from HFD B6 mice display morphologic pathologies. a The echocardiography determined left ventricular (LV) mass normalized to tibia length is larger in the HFD B6 mice when compared to the CD B6 mice ( p = 0.015, N > 8). b Similarly, the immunofluorescent determined cell size is larger in the HFD B6 mice when compared to the three other mouse groups (B6 CD versus B6 HFD, p = 0.025, B6 HFD versus MRL HFD p = 0.032, N > 3). c Representative cell sizes visualized by γ-sarcoglycan staining. d , e Echocardiography also revealed increases in left ventricle anterior wall during diastole and systole in the HFD B6 hearts compared to the CD B6 hearts (N > 8, bars indicate p

Article Snippet: The membranes were incubated at 4 °C overnight in 5 % NFDM/TBST with primary antibodies [anti-AMPK, anti-pAMPK, and PGC-1α (1:1000, Cell Signaling Technology, Danvers, MA, USA); anti-ACC, anti-pACC, fibronectin (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-HKII (1:5000, Millipore, Billerica, MA, USA); VDAC, Oxidative Phosphorylation antibody cocktail (1:1000, Abcam, Cambridge, MA, USA), Glut4 (1:500, EMD Millipore, Darmstadt, Germany), and γ-sarcoglycan (1:1000, Novocastra Labs, Buffalo Grove, IL, USA)].

Techniques: Mouse Assay, Staining

Journal: Cardiovascular Diabetology

Article Title: Successful metabolic adaptations leading to the prevention of high fat diet-induced murine cardiac remodeling

doi: 10.1186/s12933-015-0286-0

Figure Lengend Snippet: Glut4 is significantly reduced in the MRL hearts. a Representative immunoblot of Glut4 in mouse hearts. b γ-Sarcoglycan loading control. c Glut4 quantification normalized to γ-sarcoglycan (N = 9, B6 CD verses MRL CD p = 0.003, B6 HFD versus MRL HFD p = 0.001). d Representative immunofluorescence of Glut4 in the cardiac septum, original magnification ×100. e Brightness intensity of lines drawn ( white dashes in Fig. 5 d) across the plasma membranes

Article Snippet: The membranes were incubated at 4 °C overnight in 5 % NFDM/TBST with primary antibodies [anti-AMPK, anti-pAMPK, and PGC-1α (1:1000, Cell Signaling Technology, Danvers, MA, USA); anti-ACC, anti-pACC, fibronectin (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-HKII (1:5000, Millipore, Billerica, MA, USA); VDAC, Oxidative Phosphorylation antibody cocktail (1:1000, Abcam, Cambridge, MA, USA), Glut4 (1:500, EMD Millipore, Darmstadt, Germany), and γ-sarcoglycan (1:1000, Novocastra Labs, Buffalo Grove, IL, USA)].

Techniques: Immunofluorescence

Journal: Cardiovascular Diabetology

Article Title: Successful metabolic adaptations leading to the prevention of high fat diet-induced murine cardiac remodeling

doi: 10.1186/s12933-015-0286-0

Figure Lengend Snippet: MRL hearts contain reduced amounts of pAMPK and AMPK. a Representative immunoblot of the phosphorylated α subunits of AMPK in mouse hearts. b γ-Sarcoglycan loading control. c pAMPK quantification normalized to γ-sarcoglycan and the B6 CD average (N = 15, B6 CD versus MRL CD p = 0.010, B6 HFD versus MRL HFD p = 0.025). d Representative immunoblot of the α subunits of AMPK in mouse hearts. e AMPK quantification normalized to the same γ-sarcoglycan blot as above and the CD B6 average (N = 9, B6 CD versus MRL CD p = 0.005, B6 HFD versus MRL HFD p = 0.038, MRL CD versus MRL HFD p = 0.037). f pAMPK/AMPK (N = 6)

Article Snippet: The membranes were incubated at 4 °C overnight in 5 % NFDM/TBST with primary antibodies [anti-AMPK, anti-pAMPK, and PGC-1α (1:1000, Cell Signaling Technology, Danvers, MA, USA); anti-ACC, anti-pACC, fibronectin (1:500, Santa Cruz Biotechnology, Santa Cruz, CA, USA); anti-HKII (1:5000, Millipore, Billerica, MA, USA); VDAC, Oxidative Phosphorylation antibody cocktail (1:1000, Abcam, Cambridge, MA, USA), Glut4 (1:500, EMD Millipore, Darmstadt, Germany), and γ-sarcoglycan (1:1000, Novocastra Labs, Buffalo Grove, IL, USA)].

Techniques:

Journal: Molecular Therapy

Article Title: Prevention of Dystrophic Pathology in Severely Affected Dystrophin/Utrophin-deficient Mice by Morpholino-oligomer-mediated Exon-skipping

doi: 10.1038/mt.2009.248

Figure Lengend Snippet: Expression of dystrophin restores the dystrophin-associated protein complex (DAPC) to the sarcolemma in the PPMO-treated dKO mice. DAPC components α- and β-sarcoglycan, β-dystroglycan, and nNOS were detected by immunostaining in tissue cross-section of tibialis anterior (TA) muscles from PPMO-treated dKO mice (right panel), compared with C57BL6 normal mice (left panel) and untreated dKO mice (middle panel). Bar = 100 µm. dKO, double-knockout; nNOS, neuronal nitric oxide synthase; PPMO, peptide-conjugated phosphorodiamidate morpholino oligomer.

Article Snippet: DAPC protein detection was also performed using a rabbit polyclonal antibody to neuronal nitric oxide synthase and mouse monoclonal antibodies to β-dystroglycan, α- and β-sarcoglycan according to manufacturer's instructions (Novocastra).

Techniques: Expressing, Mouse Assay, Immunostaining, Double Knockout

Journal: The American Journal of Pathology

Article Title: Simultaneous Dystrophin and Dysferlin Deficiencies Associated with High-Level Expression of the Coxsackie and Adenovirus Receptor in Transgenic Mice

doi: 10.2353/ajpath.2006.060570

Figure Lengend Snippet: Immunohistochemical investigation of CAR +/+ muscle. Transverse cryostat sections from the gastrocnemius of CAR +/+ transgenic ( A , C , E , G , I ) and wild-type ( B , D , F , H , J ) mice were subjected to immunohistochemical analysis for expression of CAR ( A , B ), dystrophin ( C , D ), α-sarcoglycan ( E , F ), caveolin-3 ( G , H ), and desmin ( I , J ). In the CAR +/+ transgenic muscle ( A ) most fibers show conspicuous diffuse sarcolemmal and cytoplasmic CAR immunostaining. B: In normal control muscle, only endplates show CAR immunoreactivity ( arrows ). There is substantial reduction or even absence of immunoreactive dystrophin ( C ) and α-sarcoglycan ( E ) in the CAR homozygous mice compared with wild-type control ( D , F ). CAR +/+ transgenic muscle shows marked increase of caveolin-3 sarcolemmal immunoreactivity of all muscle fibers of varying caliber ( G ) in comparison with wild-type control ( H ). A similar marked increase in sarcolemmal and cytoplasmic desmin immunoreactivity is also seen in the CAR homozygote ( I ) when compared with normal muscle ( J ).

Article Snippet: All other antibodies were purchased commercially: anti-desmin, anti-α-sarcoglycan, and anti-β-dystroglycan were from NovoCastra (Vector Laboratories, Burlington, ON, Canada); anti-dysferlin was from Vector Laboratories; anti-caveolin-3 and anti-neuronal nitric-oxide synthase (anti-nNOS) were from Transduction Laboratories (BD Biosciences, Mississauga, ON, Canada).

Techniques: Immunohistochemistry, Transgenic Assay, Mouse Assay, Expressing, Immunostaining