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. 2013 Apr 15;24(8):1122–1133. doi: 10.1091/mbc.E12-11-0817

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© 2013 Tang et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 2: — The ndc80-NH12 mutant displays two types of mitotic abnormalities. (A) Profiles of mitotic progression. Wild-type and ndc80-NH12 cells that contained Sad1-dsRed (an SPB marker) and cen2-GFP (a GFP-tagged centromere on chromosome II) were grown at 27°C and shifted to 36°C. Mitotic cells were recorded under a fluorescence microscope, by which the distance of the two SPBs was measured and plotted against time (n = 20 for wild type and n = 40 for ndc80-NH12). Phase I is the period of initial spindle elongation (I), whereas phase II represents the duration in which the spindle length is constant (II). Phase III corresponds to anaphase B (III), when spindles elongate toward the cell tips. (B) The duration of the preanaphase stage. The duration of phases I and II was measured in four indicated strains and plotted. The data in A were used for wild type and ndc80-NH12. As for the mad2 deletion (Δmad2) and ndc80-NH12 Δmad2 strains, similar analysis as shown in A was performed (n = 13 and 12, respectively). Note that mitotic delay observed in ndc80-NH12 was abolished by the mad2 deletion. (C) Time-lapse live image analysis of sister centromere behavior. Mitotic wild-type cells (top) or ndc80-NH12 cells (bottom two rows) containing cen2-GFP (green) and Sad1-dsRed (red) cultured at 36°C were recorded (1-min intervals) and converted to kymograph. Top left, sample number (n = 55 for wild type and n = 57 for ndc80-NH12) and percentage of cells showing each phenotype (type I, 26.3%; type II, 10.5%). Type I (middle) displayed mitotic arrest during phase II, whereas type II (bottom) exhibited sister chromatid segregation errors during anaphase without mitotic delay. Scale bar, 5 μm.