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. 2018 Nov 13;8:16742. doi: 10.1038/s41598-018-34918-2

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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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(A) Size-exclusion chromatography assays. Blue: Free His6-Stl_SaPImw2 chromatogram (PK116 and PK165)_. Green: His6-DUF3113_ɸSa2mw chromatogram (PK187). Orange: Chromatogram corresponding to the interaction between His6-Stl_SaPImw2 and His6-DUF3113_ɸSa2mw. As confirmed by SDS-PAGE (top insert), PK116 belonged to His6-Stl_SaPImw2, PK187 to His6-DUF3113_ɸSa2mw and PK135 to the complex between the two proteins. (B) Size exclusion chromatography of the pull-down assays. Orange: Chromatogram corresponding to the pull-down of His6-Stl_SaPImw2 with untagged DUF3113_ɸSa2mw, in which one peak was observed for PK135, corresponding to the interaction of both proteins. Blue: Chromatogram of the pull-down of His6-Stl_SaPImw2 with untagged DUF3113_ɸSa2c. Two peaks, PK116 and PK165, corresponding to free His6-Stl_SaPImw2were observed. All peaks were analysed by SDS-PAGE. Inj1: injection of His6-Stl_SaPImw2 (32,5 KDa) and His6-DUF3113_ɸSa2mw (10.6 KDa); Inj2: injection of His6-Stl_SaPImw2; Inj3: injection of His6-DUF3113_ɸSa2mw; Inj4: injection corresponding to the pull-down of His6-Stl_SaPImw2 - untagged DUF3113_ɸSa2mw (7.1 KDa); and Inj5: injection corresponding to the of pull-down His6-Stl_SaPImw2-untagged DUF3113_ɸSa2c (7.1 KDa). A degradation product from protein His6-Stl_SaPImw2 was observed in PK165 (asterisk).