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. 2013 Jan 18;32(3):424–436. doi: 10.1038/emboj.2012.348

Figure 6.

Figure 6

The Ndc80 complex is targeted to kinetochores by two parallel pathways. (A) Kinetochore localization of Spc25 I156R mutants was reduced to ∼60% in DT40 cells. Immunofluorescence images showing the co-localization of chicken wild type or I156R mutant Spc25-GFP with CENP-T. Signal intensities of each protein were measured relative to an adjacent background signal. Bar, 10 μm. (B) Graph showing growth curves of DT40 cells in which expression of Spc25 is replaced with Spc25 I156R mutant. The doubling time of these cells was 14.2 h compared to 13.1 h for control cells. Tetracycline was added at time 0 to repress transcription of wild-type Spc25. (C) Spc25 mutants defective for CENP-T interactions require the Mis12 complex to localize to kinetochores. Images showing localization of Spc25 (I156R) in Dsn1- or CENP-T-degron cells. Dsn1 or CENP-T was degraded within 1 h after the addition of auxin (see Supplementary Figure S6).