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. 2019 Feb 12;9:1839. doi: 10.1038/s41598-018-37949-x

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© The Author(s) 2019

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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Flagellar motor switch FliG silencing reduces bacterial motility. (A) L. biflexa colonies containing wither pMaOri.dCas9 or pMaOri.dCas9sgRNAfliG were visualized under light microscope and their diameter calculated. (B) Graphic shows the average ± standard deviation of 15 colonies of each plate. For statistical analysis, diameter values displayed by cells containing dCas9 and sgRNA were compared to the ones displayed by cells containing pMaOri.dCas9 plasmid alone by Student’s two-tailed t test (*P < 0.05). (C) Different colonies were selected, grown in liquid media and cell extracts of two distinct leptospiral pools were transferred onto nitrocellulose membrane, which was probed with mouse anti-recombinant FliG (1:500) and HRP-conjugated anti-mouse IgG (1:5,000); after development with chemiluminescent substrate, the same membrane was washed and incubate with mouse anti-DnaK antiserum (1:3,000) as loading control.