Xenbase Image ID: 118756

Figure 5. Reconstitution of mitosis-specific dissociation of human 14S cohesin complexes and of PDS5 from chromatin in Xenopus egg extracts. (A) HeLa chromatin was incubated in Xenopus interphase extract. The extract was either supplemented with nondegradable cyclin B Δ90 to trigger entry into mitosis (left) or left untreated (right). At different time points either extract samples (top) or chromatin reisolated from the extract by sucrose cushion centrifugation (bottom) were analyzed by SDS-PAGE and either PhosphorImaging (top) or immunoblotting with antibodies to the indicated proteins (bottom). The cell cycle state of the extracts was analyzed by monitoring the phosphorylation-dependent electrophoretic mobility shift of 35S-labeled CDC25 and the stability of 35S-labeled cyclin B, which were added to the extracts at time zero. TOPO II, topoisomerase II; H3P, histone H3 phosphorylated on serine 10. (B) HeLa chromatin was incubated in mitotic Xenopus egg extract (xtΔ90), or in mitotic extract treated with 0.8 mM roscovitin (XΔ90+ Roscovitin), or in interphase extract (xti). Chromatin bound proteins were isolated at different time points and analyzed as in A. Image published in: Sumara I et al. (2000) © 2000 The Rockefeller University Press. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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