glut4 antibody  (Santa Cruz Biotechnology)

Journal: Antioxidants

Article Title: PIN (Protein Inhibitor of Neuronal Nitric Oxide Synthase) Modulates Glucose Uptake Through NO-Dependent and Independent Mechanisms in Rat Muscle Cells

doi: 10.3390/antiox15040436

Figure Lengend Snippet: Effect of PIN overexpression on glucose uptake and GLUT4 translocation in L6 and primary muscle cells. ( A ) Western blotting analysis of PIN overexpression after transfection with PIN cDNA (PIN) or the empty vector (C) in L6 myocytes. ( B ) Effects of PIN overexpression on glucose uptake in L6 myocytes with or without 100 nM insulin. ( C ) Effects of PIN overexpression on glucose uptake in primary myotubes with or without 100 nM insulin. ( D ) Western blotting analysis of GLUT1 expression after transfection with PIN cDNA (PIN) or the empty vector (C) in L6 myocytes. ( E ) Western blotting analysis of GLUT4 expression after transfection with PIN cDNA (PIN) or the empty vector (C) in L6 myocytes. ( F ) Colorimetric immunoassay measuring GLUT4 translocation after PIN overexpression in L6 myocytes with or without 100 nM insulin. * p < 0.05; ** p < 0.01; and *** p < 0.001.

Article Snippet: After saturation with PBS-0.4% BSA during 30 min, cells were incubated with an anti-GLUT4 antibody directed against 13 N-terminus extracellular amino acid of GLUT4 (GT42A, Alpha Diagnostic, San Antonio, TX, USA) overnight.

Techniques: Over Expression, Translocation Assay, Western Blot, Transfection, Plasmid Preparation, Expressing

Journal: Antioxidants

Article Title: PIN (Protein Inhibitor of Neuronal Nitric Oxide Synthase) Modulates Glucose Uptake Through NO-Dependent and Independent Mechanisms in Rat Muscle Cells

doi: 10.3390/antiox15040436

Figure Lengend Snippet: Effect of nNOS blockade by L-NAME on glucose uptake and GLUT4 translocation in L6 and primary muscle cells. Effects of D-NAME ( A ) and L-NAME ( B ) on glucose uptake in L6 myocytes with or without 100 nM insulin. ( C ) Effects of L-NAME on glucose uptake in primary myotubes with or without 100 nM insulin. ( D ) Colorimetric immunoassay measuring GLUT4 translocation in L6 myocytes in the presence of 10 mM L-NAME with or without 100 nM insulin. ( E ) Effect of L-NAME on nNOS catalytic activity with or without 100 nM insulin. ( F ) Effects of SNP on glucose uptake in L6 myocytes with or without 100 nM insulin. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: After saturation with PBS-0.4% BSA during 30 min, cells were incubated with an anti-GLUT4 antibody directed against 13 N-terminus extracellular amino acid of GLUT4 (GT42A, Alpha Diagnostic, San Antonio, TX, USA) overnight.

Techniques: Translocation Assay, Activity Assay

Journal: Antioxidants

Article Title: PIN (Protein Inhibitor of Neuronal Nitric Oxide Synthase) Modulates Glucose Uptake Through NO-Dependent and Independent Mechanisms in Rat Muscle Cells

doi: 10.3390/antiox15040436

Figure Lengend Snippet: Effects of PIN silencing and disruption with its interacting partners on glucose uptake in L6 muscle cells. ( A ) Western blotting analysis of PIN under-expression after transfection with PIN and control (C) siRNA. ( B ) Effects of PIN silencing on glucose uptake in L6 myocytes with or without 100 nM insulin. ( C ) Western blotting analysis of GLUT1 expression after transfection with PIN and control (C) siRNA. ( D ) Western blotting analysis of GLUT4 expression after transfection with PIN and control (C) siRNA. ( E ) Colorimetric immunoassay measuring GLUT4 translocation in L6 myocytes after PIN silencing in the presence or not of 100 nM insulin. ( F ) Western blotting analysis of nNOS expression in L6 myocytes after transfection with PIN or control (C) siRNA. ( G ) Effect of PIN silencing on nNOS catalytic activity in the absence of insulin. ( H ) Coimmunoprecipitation of nNOS with PIN in the presence of the control (peptide C) versus the inhibitory peptide (peptide I) in L6 myocytes. ( I ) Effects of the inhibitory peptide (peptide I) on glucose uptake in L6 myocytes with or without 100 nM insulin versus the irrelevant one (peptide C). ( J ) Effects of the inhibitory peptide (peptide I) on glucose uptake in primary myocytes from the insulin resistant Zucker fa/fa rat. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: After saturation with PBS-0.4% BSA during 30 min, cells were incubated with an anti-GLUT4 antibody directed against 13 N-terminus extracellular amino acid of GLUT4 (GT42A, Alpha Diagnostic, San Antonio, TX, USA) overnight.

Techniques: Disruption, Western Blot, Expressing, Transfection, Control, Translocation Assay, Activity Assay