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. 2009 Dec 1;23(23):2778–2791. doi: 10.1101/gad.541309

Figure 4.

Figure 4.

Spindle-associated Stu1 relocates to detached KTs. (A) Stu1 relocates from captured KTs to the spindle and then returns to the remaining detached KTs. Cells were processed as in C. Magnification of the boxed area is as indicated. (White arrows) Captured KTs; (red arrows) detached KTs that acquired Stu1 only in the second round. Bar, 2 μm or as shown. (B) Stu1 relocates to detached KTs in metaphase. GAL-CDC20 cells were arrested in metaphase by Cdc20 depletion, treated with nocodazole, and visualized by time-lapse microscopy. (0′) Nocodazole addition. Magnification of the boxed area is as indicated. Bar, 2 μm or as shown. (C) Stu1 relocates to KTs when the okp1-52 mutation is induced in metaphase. GAL-CDC20 cells were arrested in metaphase by Cdc20 depletion, then shifted to 37°C and analyzed at the indicated time after the temperature shift. Cells with KT-localized Stu1 were quantified (n > 100) and classified into type 1 + 2 (intact spindle) and type 3 + 4 (defective spindle). Bar, 2 μm.