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. 1999 Sep 6;146(5):941–954. doi: 10.1083/jcb.146.5.941

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© 1999 The Rockefeller University Press

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 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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graphic file with name JCB9904020.f6a.jpg — hBUBR1 kinase in metaphase Hela cells are stimulated by disruption of the spindle. Mitotic shakeoff cells that were nearly uniformly at metaphase were exposed to nocodazole for the times indicated and hBUBR1 complexes were immunoprecipitated from cell lysates and probed for autokinase activity. (A) hBUBR1 autokinase activity from each time point was quantitated with a PhosphorImager and normalized to metaphase cells (time 0). This is a valid comparison as equal amounts of hBUBR1 were used for the kinase assay. (B) Western blots of hBUBR1 immunoprecipitates probed with hBUBR1 (top) and hBUB3 (middle). Lysates obtained from the same experiment were separately probed for cyclin B expression (bottom). (C) Western blot of the insoluble chromosome fractions obtained from the experiment described above was probed for hBUBR1.

graphic file with name JCB9904020.f6bc.jpg — hBUBR1 kinase in metaphase Hela cells are stimulated by disruption of the spindle. Mitotic shakeoff cells that were nearly uniformly at metaphase were exposed to nocodazole for the times indicated and hBUBR1 complexes were immunoprecipitated from cell lysates and probed for autokinase activity. (A) hBUBR1 autokinase activity from each time point was quantitated with a PhosphorImager and normalized to metaphase cells (time 0). This is a valid comparison as equal amounts of hBUBR1 were used for the kinase assay. (B) Western blots of hBUBR1 immunoprecipitates probed with hBUBR1 (top) and hBUB3 (middle). Lysates obtained from the same experiment were separately probed for cyclin B expression (bottom). (C) Western blot of the insoluble chromosome fractions obtained from the experiment described above was probed for hBUBR1.