Fig. 5
From: RNAs coordinate nuclear envelope assembly and DNA replication through ELYS recruitment to chromatin
RNAs are required for nuclear pore complex assembly and nuclear import. a Representative images of immunofluorescence analysis of nuclei fixed after incubation in mock- or RNase A-treated X. laevis egg extracts for 20 min. The mouse monoclonal antibody 414 (mAb414) recognises several nucleoporins (NUP358, 214, 153, 62); DNA was stained with DAPI (blue). Scale bar, 10 μm. b Western blot analysis of nuclear extracts after incubation in mock- or RNA-depleted egg extracts for the indicated time. c Sperm nuclei were incubated in mock- or RNA-depleted egg extracts. At the indicated time points, chromatin was isolated and immunoblotted with the indicated antibodies. d Immunofluorescence analysis of ELYS (green) and NUPs (red) expression in a representative population of nuclei assembled in mock- or RNase A-treated extracts for 60 min. Images were acquired with an ApoTome microscope. Scale bar, 15 μm. e Nuclear swelling and DNA decompaction are restored when nuclei are first incubated in RNase A-treated extracts and then transferred to mock-treated extracts (as explained in Fig. 3c). After transfer into the second extract, nuclei were fixed and stained with DAPI and DiOC6 to monitor decompaction and nuclear swelling. Scale bar, 10 µm
