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. 2022 Mar 28;13:1641. doi: 10.1038/s41467-022-29187-7

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2022

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. 1 — a Sequences of double-stranded (ds) nucleic acids used for S9.6 characterizations and locations of FAM labels. b Schematic representation and denaturing SDS-PAGE analysis of S9.6 Fab in oxidizing and reducing conditions. SDS-PAGE analysis was performed twice. HC heavy chain, LC light chain. c SEC-MALS profiles of free S9.6 Fab (black), free DNA-RNA hybrid (magenta), free dsRNA (red), and free dsDNA (blue) and stoichiometric (1:1) mixtures of S9.6 Fab with hybrids (green), dsRNA (salmon), and dsDNA (light blue). SEC-MALS-derived molecular weights are indicated. d Binding affinity measurements of S9.6 Fab with nucleic acids in a by fluorescence polarization titration. Apparent binding constants (K_ds) are indicated. Values are mean ± s.d. n = 3 biologically independent samples. ΔFP: changes in fluorescence polarization, in mP units. ND: not determined. e Competition experiments of fluorescently labeled DNA-RNA hybrids bound to S9.6 with increasing amounts of unlabeled nucleic acids, colored as in d. Apparent IC₅₀s are indicated. Values are mean ± s.d. n = 3 biologically independent samples. f Isothermal titration calorimetry (ITC) profile of S9.6 binding to DNA-RNA hybrid. The measured binding constant is indicated. Values are mean ± s.d. n = 2 biologically independent samples. g ITC profile of S9.6 binding to dsRNA. Source data are provided as a Source Data file.