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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 4. — TOGL1 and ML localize the Stu1 dimer to metaphase KTs but not to anaphase KTs. (A and B) The Stu1–KT interaction in metaphase requires TOGL1, ML, and dimerization. Cells were arrested in metaphase by Cdc20 depletion for 3 h and analyzed by ChIP. (A) The presence of CEN3 DNA and two flanking DNA regions (CHIII-R and CHIII-L) was detected by triplex PCR. The input was 0.1% of the immunoprecipitation (IP). (B) Quantitative ChIP. The mean (3.5-fold) enrichment of CEN3 DNA over a control DNA (PHO5) observed for the WT was set to 100%. Error bars represent the SDs of three PCR experiments. (C) Stu1 is absent from anaphase KTs. cdc15-1 cells were arrested in anaphase by incubation at 37°C and analyzed by quantitative ChIP as in B. The result obtained for cells arrested in metaphase (set to 100%) is shown as a comparison.