Figure 2.

MAD-1 accumulation requires kinetochore-localized BUB-1. (A) The fluorescence intensity of GFP::MAD-1 at unattached kinetochores in monopolar spindles was quantified and expressed as a fraction of that in controls. n is the number of embryos; error bars are the 95% confidence interval of the mean. Ctrl, control. (B) Schematic showing the deleted/mutated regions in the indicated KNL-1 mutants. Graph plots the ratio of BUB-1::GFP to KNL-1::mCh for each KNL-1 variant normalized by dividing by the same ratio for WT KNL-1::mCh. Endogenous KNL-1 was depleted in all conditions. n is number of embryos analyzed. Error bars are the 95% confidence interval of the mean. See also Fig. S1 B. (C and D) MAD-1 and BUB-1 targeting to unattached kinetochores in strains expressing the indicated KNL-1 variants after depletion of ZYG-1 and endogenous KNL-1. n is the number of embryos analyzed. Graph plots the ratio of GFP::MAD-1 (in a mad-1Δ background) or BUB-1::GFP to KNL-1::mCh at kinetochores of monopolar spindles in the indicated strains. Error bars are the 95% confidence interval of the mean. Bars: (A and C) 5 µm; (B) 2 µm.

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