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Figure 6


Fig. 6. Mitch localizes to the kinetochore during mitosis. (A) Wild-type (top row) and mitch-mutant (bottom row) brains were stained with anti-Mitch antibodies (red). In wild-type, Mitch localizes to kinetochores from prometaphase through late anaphase and likewise in chromosome spreads of cells arrested in metaphase by colchicine (c-metaphase). The kinetochore staining of anti-Mitch antibodies is absent in mitotic figures from mitch mutants. (L-R on the bottom row: mitch1, mitch2/Df, mitch2/Df, mitch1). (B) Epitope-tagged Mitch. Western blot (left panel) of Drosophila tissue culture cells non-transfected (NT) or transfected with mitch-V5, and probed with anti-V5 antibody, identifying the 30 kd Mitch-V5 protein. (Right panels) Indirect immunofluorescence of cells incubated with anti-V5 antibody. Mitch-V5 was absent in non-transfected cells (NT), but localized to the kinetochore in transfected cells (here seen in c-metaphase). Bar, 5 µm.





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