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Fig. 2. Immunolocalization of Mad2, Bub1, Bub3 and BubR1 during progression through mitosis in Drosophila S2 cells. Cells were stained for DNA (blue) and antibodies against Mad2, Bub1, Bub3 or BubR1 (red). {alpha}-tubulin staining (green) was used to identify cells in prophase. Note that only Bub3 and Bub1 show kinetochore staining in prophase. All spindle checkpoint proteins show strong accumulation at kinetochores in prometaphase, weak or no staining in metaphase and are absent in anaphase. Staining of the spindle for Mad2 and Bub1 is observed during metaphase, and weak staining of centrosomes (arrowheads) is observed for all proteins at anaphase. Scale bar: 5 µm.





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