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. 2009 Jan 1;20(1):10–20. doi: 10.1091/mbc.E08-03-0324

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© 2008 by The American Society for Cell Biology

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Figure 5. — Kinetochore targeting of the N-terminal fragment of Mps1 requires phosphorylation at T12 and S15. (A) Kinetochore localization of YFP-Mps1, YFP-Mps1^1-301, and YFP-Mps1^{1-301T12S15→AA} in prometaphase-arrested cells induced by nocodazole treatment (0.1 μg/ml). The kinetochores and chromosomes are identified by staining with human CREST antisera and DAPI, respectively. (B) Quantitation of fluorescent density of YFP-Mps1 and related mutants on the kinetochores of prometaphase cells. More than 52 kinetochores in at least five randomly selected cells in each cell line were analyzed. YFP-Mps1^{1-301T12S15→AA} cannot relocalize to kinetochores in prometaphase cells. (C) Immunoblotting analysis of expression of YFP-Mps1^1-301, and YFP-Mps1^{1-301T12S15→AA} in SW480 cells.