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. 2021 Sep 20;12:5545. doi: 10.1038/s41467-021-25830-x

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© The Author(s) 2021

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. 3 — a Average normalized Cy3-43mer fluorescence profiles plotted as a function of time. The arrow indicates the components of the two syringes rapidly mixed at the 0 s time point in a stopped flow instrument. RAD-51-ssDNA filaments pre-formed with RAD-51 (1 μM) and Int(21)-Cy3 labelled 43mer ssDNA (15 nM) and indicated concentrations of RFS-1/RIP-1 were mixed for 10 min in the presence of ADP. The mixture was then challenged with 100-fold excess of unlabelled 43mer and analyzed (n = 5–8). b Graph of RFS-1/RIP-1 concentration-dependence change of Cy3 fluorescence from (a) (mean; errors: s.d.). c Representative EMSA gel (n = 4) demonstrating destabilization of RAD-51-ssDNA filaments in the presence of ADP. RAD-51 (1 μM) and RFS-1/RIP-1 (0.5 μM) were pre-incubated before addition of 5′-FITC-61mer ssDNA (10 nM) for 10 min and then challenged with increasing amounts of unlabelled 61mer ssDNA for a further 10 min. Protein-DNA complexes were crosslinked and resolved in agarose gels. d The amount of unbound 5′-FITC-61mer ssDNA from (c) (n = 4) was quantified. The average percentage of unbound 5′-FITC-61mer ssDNA was plotted as a function of the relative excess concentration of unlabelled 61mer over 5′-FITC-61mer ssDNA (mean; errors: s.d.). Source data are provided as a Source Data file.