Fig. 6. The interphase function is not blocked 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. After 30 minutes, biotinylated-dUTP was added. After 90 minutes, replication foci were visualized with Alexa-488-labeled streptavidin, and chromatin visualized with DAPI. Control samples in the lower row were treated with aphidicolin. (B) Nuclei were assembled as in A. After 90 minutes, a fluorescently labeled import substrate (upper row) or a control substrate (lower row) were added. The reaction was stopped after 30 additional minutes by fixation and isolated. Chromatin is stained with DAPI (blue) and membranes with DiIC18 (red). (C) Nuclei were assembled as in A. After 90 minutes, a biotinylated nondegradable cyclin B mutant protein was added. The reaction was stopped after 30 additional minutes by fixation and isolated. Biotinylated cyclin B was visualized with fluorescently labeled streptavidin (green) and NPCs by means of mAb414 staining (red). Bars, 10 µm.