JCS03235 Supplementary Material

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  Fig. S1. Mitotic spindles in ncd and sub mutants stained for DNA (blue) and microtubules (green). (A) Relatively normal bipolar spindle in ncd mutant metaphase. The ncd spindles were usually bipolar with few (see Table 1, 18.1%) organizational defects. The only significant problem was that these mutants exhibited indications of a defect in spindle pole maintenance resulting in a split in the spindle, giving the appearance of two separate spindles connected to the same pole. Like sub mutants, however, ncd mutants exhibited a high frequency of lagging chromosomes at anaphase (Table 2). In addition, the ncd mutant brains had a low metaphase to anaphase ratio (Table 2). Together with the mitotic index data, we suggest that ncd mutant mitotic cells progress rapidly through metaphase, often resulting in chromosome alignment defects and errors in segregation. (B) The sub;ncd double mutant exhibited cytological phenotypes that were mostly additive and a combination of the two single mutant phenotypes, exhibiting, for example, asymmetric half spindles. These phenotypes included spindle pole defects, a low metaphase to anaphase ratio, disorganized metaphase spindles, asymmetric half spindles and lagging anaphase chromosomes (see Table 2). In less-frequent cases (C) (C’, DNA channel only), the sub;ncd double mutant appeared to be entering anaphase before the spindle was assembled. Overall, however, the double mutant spindle phenotype was not more severe than the single mutant phenotypes. ncd and sub do not appear to have redundant functions in spindle assembly. Bar, 5 μm.