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. 2002 Jul 8;158(1):23–29. doi: 10.1083/jcb.200202053

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Copyright © 2002, The Rockefeller University Press

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Figure 5. — Fluorescence loss in photobleaching demonstrates exchange of topoIIα between chromosomal and cytoplasmic pools in mitotic cells. (A) Exchange of topoIIα occurs between the chromosomes and the cytoplasm of mitotic cells. A late prometaphase cell outlined by the solid line was targeted for photobleaching in the cytoplasm away from the chromosomes indicated (dotted circle). The region was photobleached every 20 s and images were captured every 2 min. Repeated photobleaching of the cytoplasm results in depletion of fluorescence in the chromosomes. The fluorescence intensity of the chromosomes was reduced by 79%, whereas that of the control interphase nucleus (arrow) in the adjacent interphase cell was reduced only 8%. (B) Exchange of topoIIα occurs through the cytoplasm among chromosomes. A cell treated with 0.1 μg/ml nocodazole was repeatedly bleached in the lower region of the mass of chromosomes (dotted circle). Fluorescence intensity of the isolated chromosome at top (closed arrows) diminished by 85%, whereas the intensity of the nucleus in the neighboring cell (open arrow) decreased by only 5% over the course of the observations (Video 2, available at http://www.jcb.org/cgi/content/full/jcb.200202053/DC1). Bars, 10 μm.