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. 2018 Sep 3;217(9):3019–3029. doi: 10.1083/jcb.201707081

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© 2018 Wynne and Vallee

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

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Figure 4. — Role of Nde1 phosphorylation in mitotic progression. (A–F) Duration of mitotic events for individual H2B-RFP HeLa cells expressing GFP or GFP-tagged WT Nde1, Nde1-3E, Nde1-3A, Nde1-del-dynein, or Nde1-del-Lis1. (G) Duration of prometaphase (“unaligned”) and metaphase (“aligned”) is shown and replotted as the complete time from NEBD to anaphase onset. Mean ± SEM for each condition is shown. Mann–Whitney statistical tests revealed a significant increase in the time from NEBD to anaphase onset for cells expressing GFP Nde1-3A, GFP Nde1-del-dynein, and GFP Nde1-del-Lis1 relative to GFP alone control. (H–N) Same as above, but in H2B-RFP–expressing HeLa cells treated with control or Nde1 siRNAs for 48 h and then rescued with GFP, WT Nde1, Nde1-3E, Nde1-3A, Nde1-del-dynein, or Nde1-del-Lis1 for 24 h before live-cell imaging. (O) Data replotted as complete time from NEBD to anaphase onset. Mann–Whitney statistical tests were performed to compare Nde1 siRNA to each GFP Nde1 rescue condition. All of the Nde1 constructs to some extent rescued effects of Nde1RNAi, whereas Nde1-del-dynein showed no such effect. Mean ± SEM for each condition is shown. **, P < 0.01; *, P < 0.05; ****, P < 0.0001.