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. 2017 Mar 7;8:14678. doi: 10.1038/ncomms14678

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Copyright © 2017, The Author(s)

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(a) Schematic of the experiment. CENP-A was knocked down for two days prior to conducting a double thymidine block and release. Twelve hours after the second release, cells were treated with Mg132 or Mg132 and Monastrol for one hour. (b) Immunofluorescence analysis showing multipolar spindles formed in cells treated with CENP-A shRNA are rescued by treating cells with the Eg5 inhibitor Monastrol. Scale bar, 10 μm. (c) Graph showing fractions of monopolar, bipolar and multipolar cells in cells treated with Eg5 inhibitor after CENP-A shRNA treatment. (d) Graph showing fractions of monopolar, bipolar and multipolar cells in cells treated with various concentrations of Eg5 inhibitor after CENP-A shRNA treatment and replacement with CENP-A wild-type and methylation mutant. The experiment was done in duplicate and ∼100 mitotic cells were counted in each condition. Error bars indicate s.d. Scale bars, 10 μm.