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. 2019 Jul 22;10:3274. doi: 10.1038/s41467-019-11104-0

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — At the activation step of DNA replication, OGRE/G4 elements compete for dsDNA but not for ssDNA replication. a Schematic representation of the replication kinetics of sperm nuclei in X. laevis low-speed egg extracts (LSE) in the presence of competing double-stranded (ds) oligonucleotides. LSEs were pre-incubated with competing oligonucleotides, sonicated salmon sperm DNA, or ultrapure H2O (mock) at 22 °C for 5 min. b Average DNA replication efficiency (mean + SD) of LSEs pre-incubated with competing oligonucleotides or controls (n = 6 for mock/salmon sperm DNA pre-incubated extracts, n = 3 for mock/random oligonucleotides). Total incubation time was 2 h. P values were obtained using the two-tailed Student’s t test; ****p < 0.0001, p > 0.05. c Replication kinetics of ssM13 complementary DNA strand synthesis in X. laevis high-speed egg extracts (HSE) pre-incubated with the indicated competing oligonucleotides or H2O (mock). d Competition by OGRE/G4 oligonucleotides does not affect pre-RC formation. Sperm nuclei were added to HSEs incubated with H2O (mock), sonicated salmon sperm DNA (S.Sperm), or the indicated oligonucleotides. Chromatin was isolated and immunoblotted with the indicated antibodies. Histone H2B level was used as loading control. e, f Competition by OGRE/G4 oligonucleotides affects DNA replication activation. Time-course analysis of replication initiation factor recruitment to chromatin after addition of sperm nuclei to LSEs pre-incubated with H2O (mock), sonicated salmon sperm DNA (S. sperm), or competing oligonucleotides. At the indicated time points, chromatin was isolated and immunoblotted with the indicated antibodies; ns not significant