A Kinase Anchoring Protein 9 Is a Novel Myosin VI Binding Partner That Links Myosin VI with the PKA Pathway in Myogenic Cells

Figure 4

Validating the AKAP9 and MVI interaction. (a) Coimmunoprecipitation of AKAP9 (left panel) and DOCK7 (right panel) with anti-MVI antibody from HEK 293 cells overexpressing GFP-tagged full length human MVI heavy chain. Left lanes: cell lysate (lysate), middle lanes: samples precipitated with a normal rabbit serum (IgG-IP), and right lanes: αMVI-IP. Samples precipitated with anti-MVI antibody were probed with either anti-AKAP9 or anti-DOCK7 antibodies as marked on the figure. PLA assay probing MVI and AKAP9 interactions (in red) in undifferentiated myoblasts (b) and in myotubes (c). In blue, nuclei stained with DAPI. (c) MVI-AKAP9 possible interactions are also seen close to bungarotoxin- (BTX-) stained acetylcholine-rich clusters (in green) marked by arrowheads. The lower panels in (b) and (c) are ~3x magnifications of the areas marked in (b) and (c). Images in (b) and (c) of the cell central sections (z = 0.3μm) were obtained with a Leica confocal microscope. Bars: 20μm.

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