Figure 4.

Normal nuclear assembly and function in the absence of RanBP1 and RCC1. (A) Nuclear assembly occurs normally in the absence of both RanBP1 and RCC1. Control (top), codepleted (middle), and mock-depleted (bottom) cytosol were used for a standard nuclear assembly assays. After 30, 60, and 120 min of incubation, nuclei were stained with Hoechst 33258 DNA dye. Bar, 2.6 × 10⁻⁶ m. (B) DNA replication occurs normally in the absence of both RanBP1 and RCC1. Nuclei assembled as in A were allowed to carry out DNA replication in the presence of [α-³²P]dCTP. At 90 min and 150 min, DNA samples from control (row 1), codepleted (row 2), and mock-depleted (row 3) reactions were taken for analysis of ³²P incorporation on agarose gels. The samples were treated as previously described (Smythe and Newport, 1991), and replication was quantified using a Molecular Dynamics Phosphorimager.