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Figure 5


Fig. 5. NEBD is inhibited by Fab fragments against the C-terminal domain of gp210. (A) Nuclei were assembled in the presence of either control Fab fragments against the lumenal domain of gp210 or inhibitory Fab fragments against the C-terminal domain of gp210, respectively. After two hours, a twofold volume of mitotic extract was added and the samples incubated for another 2 hours. Membranes were stained by DiIC18, chromatin with DAPI and samples analyzed by confocal microscopy. (B) An experiment was performed as in A except that, instead of mitotic extracts, cyclin B {Delta}90 was added to drive the system into mitosis. (C) An experiment was performed as in A except that the Fab fragments against the indicated nucleoporins were added after 110 minutes and mitotic extract after 120 minutes. (D) An experiment was performed as in A except that samples were analyzed using antibody against NUP153 (green) and mAb414 (red). Chromatin was stained with DAPI (blue in overlays). (E) An experiment was performed as in A except that samples were analyzed using an antibody against phospho-histone H3 (green) and the monoclonal antibody mAb414 (red). Chromatin was stained with DAPI (blue in upper row). Bars, 10 µm.





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