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. 2011 Dec 1;22(23):4486–4502. doi: 10.1091/mbc.E11-07-0608

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© 2011 Buttrick 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 3: — Nsk1 loading occurs at anaphase onset. (A) Log-phase cultures of nsk1-gfp sid4-tdtomato cells were grown on agar pads at 30°C and analyzed by fluorescence microscopy. A series of images of a mitotic cell at 1-min intervals is depicted. The time at which Nsk1 appears at the kinetochore–SPB junction (open arrows) and late anaphase B spindle (closed arrow) is highlighted. Scale bars: 2 μm. (B) Graph of spindle length against time for multiple nsk1-gfp sid4-tdtomato cells. Spindle length is the distance between two Sid4-tdtomato dots. Time zero corresponds to the appearance of Nsk1 at the pole. (C & D) Log-phase cultures of nsk1-gfp sid4-tdtomato (C) or nsk1-gfp cnp1-mcherry (D) cells were grown on agar pads at 30°C and analyzed by fluorescence microscopy. A series of images from two representative cells at 30 s intervals are depicted. Note that in each case Nsk1 appears before the end of anaphase A. Time 0 corresponds to the appearance of Nsk1 at the poles.