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Figure 4. Smarcal1-Dependent Regulation of RVFs(A) EM micrograph showing RVF intermediate isolated from extracts treated with H-APH (1.5 mM). The arrow indicates a double-stranded reversed branch. The arrowhead indicates the single-stranded tail of the reversed branch. The inset shows a high magnification of the RVF junction.(B) Chromatin binding of the indicated factors at the indicated times following buffer or H-APH supplementation. Buffer and H-APH were added to egg extract 45 min after sperm nuclei. NS, absence of sperm nuclei.(C) RVF frequency in extracts treated as indicated. Mean values ± SEM (n = 3) are shown. ∗,∗∗,∗∗∗p < 0.01, obtained by unpaired t test between the marked couples.(D) Scheme showing the assay to quantify RVFs with ssDNA tails (STAR Methods).(E) ELISA detection of BrdU in nascent ssDNA in nuclei incubated in extracts treated as shown. Where indicated, extracts were supplemented with 5 ng/μL recombinant human Smarcal1WT or catalytically dead Smarcal1HD. Mean intensity values ± SD (n = 3) are shown.See also Figure S6. Image published in: Kolinjivadi AM et al. (2017) © 2017 The Author(s). This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives license Larger Image Printer Friendly View

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