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. 2009 Jan 12;184(1):21–29. doi: 10.1083/jcb.200804098

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© 2009 Scott et al.

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.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 2. — Xpo1p-dependent targeting of Mad1p to kinetochores. (A) MAD1-GFP and MTW1-RFP in nup60Δ strains containing an otherwise WT genotype (Y3057) or the mutant alleles kap95-14 (Y3161), msn5Δ (Y3116), xpo1-T539C (Y3156), or prp20-7 (Y3091) were examined under the indicated conditions of temperature or LMB addition and after nocodazole addition. The localization of Mad1-GFP and the kinetochore protein Mtw1-RFP was examined by confocal microscopy. Note that as described previously (Gillett et al., 2004), some nocodazole-treated cells contain two kinetochore foci, with one lacking attached microtubules and being capable of binding Mad1p. Arrows point to overlapping Mad1-GFP and Mtw1-RFP foci. Arrowheads point to kinetochores devoid of Mad1-GFP. (B) Quantification of Mad1-GFP and Mtw1-RFP colocalization from the experiments presented in A. The results of three experiments were combined, and SD (error bars) is shown in each case. Bars, 2 μm.