Fig. 6. Co-localization of ß-dystrobrevin and ubiquitous kinesin heavy chain in transfected COS-7 cells. The sub-cellular distribution of ß-dystrobrevin and Kif5B was determined by transfecting COS-7 cells with pCMV-HA/ß-DB (A) and pCB6/Kif5B (B). ß-Dystrobrevin is located in intensely staining punctae throughout the cell, more concentrated around the nucleus (A), whereas kinesin immunoreactivity appears reticular within the COS cell cytoplasm (B). Co-transfection of ß-dystrobrevin and Kif5B resulted in a tubulo-vesicular pattern of the two co-expressed proteins, ß-dystrobrevin (C) and kinesin (D), with a significant co-localization (E). Nocodazole treatment of co-transfected cells resulted in a dramatic redistribution of ß-dystrobrevin (F) and kinesin (G) into vesicular aggregates, with precise co-localization (H). ß-Dystrobrevin was detected using the polyclonal ß-dystrobrevin antibody, followed by a goat anti-rabbit IgG secondary antibody conjugated with TRITC (A,C,F). Kinesin was detected using a monoclonal kinesin antibody (Clone H2), followed by a goat anti-mouse IgG secondary antibody conjugated with FITC (B,D,G). E and H are the merged images from each experiment: co-localization areas appear yellow. Scale bar, 10 µm.

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