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

Figure 7.

Figure 7.

Mps1 kinase activity is required for its kinetochore targeting and recruitment of Mad2 upon activation of the mitotic spindle checkpoint in SW480 cells. (A) Kinetochore localization of the N-terminal fragment of Mps1 requires endogenous Mps1. SW480 cells stably expressing YFP-Mps1R1-301 were transfected with control or Mps1 siRNA. Localization of YFP-Mps1R1-301 was determined by immunofluorescence microscopy as described in text. B is the same as A except kinase-deficient Mps1 mutant (YFP-Mps1R-KD) was used. (C) Mps1 kinase activity is required for kinetochore recruitment of Mad2. SW480 cells stably expressing YFP-Mps1R-KD were transfected with control or Mps1 siRNA. Cells were fixed and stained with an antibody against Mad2 and with DAPI. (D) Quantitation of fluorescent density of kinetochores labeled by YFP-Mps1 and related mutants in control or Mps1 siRNA treated prometaphase cells. At least 42 kinetochores from prometaphase cells in each cell line were scored and differences between control and Mps1 siRNA treated samples from YFP-Mps1R-KD or YFP-Mps1R1-301 are statistically significant (p < 0.05).