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. 2018 Sep 11;8:13576. doi: 10.1038/s41598-018-31851-2

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© The Author(s) 2018

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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In vivo identification of the TorE-TorC complex. (a) hrCN-gels, first dimension. The strains were growth in anaerobiosis. The fluorescent labelled proteins were solubilized from the membrane extracts of the various recombinant strains using DDM (2%, v/v). The solubilized fractions were loaded on hrCN gel (3–14%) and in-gel detection of fluorescent labelled proteins and complexes was performed by scanning. Recombinant strains ΔtorE/pGFP-TorE (TorE) and ΔtorC/pGFP-TorC (TorC) were grown in rich medium supplemented with TMAO to induce the other components of the Tor system (+). The strain ΔtorEC carrying either pGFP-TorE-mCherry-TorC (TorE-TorC) or pmCherry-NapE;GFP-NapC (NapE-NapC) were grown in rich medium. Separated gels are represented with their respective molecular weight markers. White lines separate portions of the same initial gel (see Figure S3). (b) SDS-Gel, second dimension. A band of hrCN gel corresponding TorE-TorC was excised and after treatment with SDS and β-mercaptoethanol was submitted to SDS-12%-PAGE. The direction of the migration is indicated by arrows. (a,b) Fluorescence was revealed by scanning. Complexes TorE-TorC and NapE-NapC are indicated by arrows. M is for molecular weight markers. The pictures are representative of three independent experiments.