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Figure 4.

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Stu2p can destabilize cytoplasmic microtubules in vivo. Wild-type or stu2–10 cells were HU arrested at the permissive temperature, shifted to the restrictive temperature, and the cytoplasmic microtubule length was determined by indirect immunofluorescence and three dimensional tracking. (A) FACS® profiles of cycling wild-type or stu2–10 cells at 25°C (top) that were arrested at 25°C with HU before (middle) or after shifting the cells to 37°C (bottom). (B) Projected indirect antitubulin immunofluorescence of wild-type or stu2–10 cells arrested at 25°C with HU before (top) or after shifting the cells to 37°C (bottom). Green, microtubules; blue, DNA. Bar, 5 μm. (C) Projected indirect antitubulin immunofluorescence of wild-type (left) or stu2–10 cells (right) arrested at 25°C with HU after shifting the cells to 37°C. Close-ups of four representative cells. Green, microtubules; blue, DNA. Bar, 5 μm. (D) Length distribution (in 1-μm intervals) of cytoplasmic microtubules of wild-type and stu2–10 cells arrested at 25°C with HU before (top) or after shifting the cells to 37°C (bottom). Green, wild-type microtubules; red, stu2–10 microtubules. (E) Average length of cytoplasmic microtubules of wild-type or stu2–10 cells arrested at 25°C with HU before and after shifting the cells to 37°C. Error bars represent SEM (P < 0.05).

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