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Fig. 3. Tension across bi-oriented kinetochore pairs of S2 metaphase cells is reduced after taxol treatment. (A) Cells in prophase or metaphase were stained for CID (green), -tubulin (red) and DNA (blue). The distance between sister kinetochores in taxol-treated metaphase cells (right panel) is shorter than in control metaphase cells (middle panel) and closer to the distance in prophase cells (left panel). Boxes of identical size are used to compare interkinetochore distances between control and taxol treated cells. Tubulin staining shows that kinetochores still retain their microtubule fibers after taxol treatment. (B) Histogram showing the variation in distances between sister kinetochores in prophase, metaphase and taxol-treated metaphase cells. Interkinetochore distances in prophase cells (range: 0.48-0.92 mm). After sister kinetochores become bioriented poleward forces stretch centromeric heterochromatin and generate tension across kinetochore pairs (range: 0.88-1.59 mm). Taxol relieves tension by interfering with microtubule dynamics causing the distance between sister kinetochores to decrease (range: 0.59-1.00 mm). (C) S2 cells showing CID (red), 3F3/2 (green) and DNA (blue). After taxol treatment, all kinetochores in metaphase cells exhibit strong 3F3/2 labelling because of lack of tension (right panel). In a control metaphase cell, 3F3/2 labelling at the kinetochores is hardly detectable because of tension-sensitive dephosphorylation (left panel). Scale bars: 5 µm.

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