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. 2020 Jun 2;10:9002. doi: 10.1038/s41598-020-66050-5

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© The Author(s) 2020

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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dbToxRp binds to ToxSp 2–4 fold more than ToxRp and the binding is not effected by bile salts. (a) Left top panel a colloidal blue stained gel excerpt showing the changes in the amount of dbToxRp pulled down when the domain was treated with buffer versus 2 mM CDC. Left bottom panel a colloidal blue stained gel excerpt showing the changes in ToxRp pulled down when the domain was treated with buffer or 2 mM CDC. Lanes 1–3 serial dilutions of the CDC treated pull down and 4–6 serial dilutions of the pull down with buffer. M is the lane with the marker. Right panel is a graph showing the relative amount of the domains pulled down by the compounds versus buffer. Each trial is represented as well as the mean and SD, n = 3. (b) Left panel gel of the periplasmic domains pulled down from the above experiments to determine the relative amount of ToxRp pulled down normalized to dbToxRp. Graph of the results normalized to the pull down performed with dbToxRp + 2 mM CDC (The intensity of dbToxRp with 2 mM CDC 1x dilution band was set to one in each trial). Each trial is represented as well as the mean and SD, n = 3.