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. 2014 May 26;205(4):555–571. doi: 10.1083/jcb.201310018

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© 2014 Funk et al.

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Figure 7. — Laterally bound Stu1 cross-links MTs. (A) Stu1 spindle localization in metaphase is independent of Ase1. Cells were arrested in metaphase by Cdc20 depletion for 2 h. (B) Stu1 cross-links MTs in vitro. As illustrated, stabilized MTs were immobilized, incubated with the indicated Stu1 constructs, and subsequently incubated with mobile MTs. Colocalization of immobile and mobile MTs was quantified. Error bars represent the SDs of two experiments. n > 100. (C) Overexpression (OE) of Stu1 arrests cells with metaphase-like spindles independent of the spindle assembly checkpoint. Yellow lines indicate the outline of the cells. Overexpression of the indicated Stu1 constructs was started after G1 release. The distribution of spindle lengths for cells with large buds (>2/3 of mother) was determined. Lines indicate the cutoff for metaphase spindles. n > 100 for WT, and n > 45 for Δmad2 cells. AMCA, aminomethylcoumarin acetate. Bars, 2 µm.