Fig. 1
From: RNAs coordinate nuclear envelope assembly and DNA replication through ELYS recruitment to chromatin
Incubation with RNase A inhibits DNA replication but not pre-RC assembly. a Replication kinetics of LSEs incubated with RNase A before (pretreat), at the same time (0 min), and 10 or 15 min after addition of sperm nuclei. b M13 ssDNA replication is not affected by LSE pre-incubation with RNase A, c Scheme illustrating the steps leading to the assembly of the complexes involved in the licensing reaction (green) and in DNA replication activation (red). The blue circle highlights reactions occurring in the nucleus. d Sperm nuclei incubated in mock or RNase A-treated LSE for 60 min were fixed in the presence of DiOC6 (green) and DAPI (blue). Scale bar, 17 μm. The nuclear size of 125 nuclei per condition was measured based on the DAPI staining area (right panel). Red bar, mean value. A two-tailed Student’s t-test was performed to determine the p-values. e Histones from sperm nuclei were isolated by acid extraction before and 5 min after incubation in mock or RNase A-treated extracts. DNA-bound proteins were then analysed by gel electrophoresis and silver staining. f Time-course analysis of pre-RC assembly in HSE (chromatin purification followed by western blotting with specific antibodies)
