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Image Search Results
Journal: Molecular Biology of the Cell
Article Title: Caveolin-3 Promotes Nicotinic Acetylcholine Receptor Clustering and Regulates Neuromuscular Junction Activity
doi: 10.1091/mbc.E09-05-0381
Figure Lengend Snippet: Caveolin-3 expression promotes activation of MuSK and Rac-1 after agrin treatment. (A) Ras G12V -transformed 3T3 cells were transiently cotransfected with caveolin-3 and each of the following constructs: pCMV-HA, pCMV-WT-MuSK-HA, pCMV-mutant A-MuSK-HA, and pCMV-mutant B-MuSK-HA. Forty eight hours after transfection, cell lysates were immunoprecipitated with an antibody probe specific for caveolin-3, and immunoprecipitates were subjected to immunoblotting analysis using anti-HA IgGs. Total expression of Cav-3 and HA-tag–expressing constructs are shown in the two bottom panels. (B–D) Myogenic precursor cells were derived from wild-type and caveolin-3 null mice, differentiated for 7 d, and treated with agrin (10 ng/ml) for 15 min, 1 h, or 4 h. Untreated cells were used as controls. (B) Cell lysates were immunoprecipitated using an antibody probe specific for MuSK, and immunoprecipitates were subjected to immunoblotting analysis using either anti-phosphotyrosine (P-Tyr) or anti-caveolin-3 IgGs. Total MuSK, caveolin-3, and phosphorylated JNK expression are shown in the bottom panel. (C) Cell lysates were immunoprecipitated using GST fused to the PBD of Pak1, and immunoprecipitates were subjected to immunoblotting analysis using anti-Rac-1 IgGs. Total Rac-1 expression is shown in the bottom panel. (D) Cell lysates were immunoprecipitated using an antibody probe specific for caveolin-3, and immunoprecipitates were subjected to immunoblotting using anti-Rac-1 IgGs. Total caveolin-3 and Rac-1 expression are shown in the bottom panel.
Article Snippet:
Techniques: Expressing, Activation Assay, Transformation Assay, Construct, Mutagenesis, Transfection, Immunoprecipitation, Western Blot, Derivative Assay