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. 2019 Aug 2;9:11216. doi: 10.1038/s41598-019-46038-6

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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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Motility of threonine mutants. (A) Motility on soft-agar plates of the threonine mutants. Overnight cultures were inoculated in soft-agar plates (0.25% agar) and incubated at 30 °C for 5 h. (B) Motility fraction of threonine mutants in the liquid medium. After 4 h of the second culture, motility was examined by light microscopy. Squares, diamonds, and circles indicate data of three independent experiments and the white bars are the total data. Numbers of bacteria examined in total are 482, 313, 390, 351, 378, 369, 291, and 350 for Vector, WT, PomA-T5A, PomA-T158A, PomA-T185A, PomA-T186A, PomB-T21A, and PomB-D24N, respectively. (C) Swimming speed of threonine mutants in the liquid medium. Thick horizontal lines are the average swimming speed. Number of bacteria examined in total is 125. (D) Expression level of stator proteins in threonine mutants. After growth for 4 h, cells were collected. The proteins were separated by SDS-PAGE and the gels were subjected to Western blotting using anti-PomA or PomB antibody. The regions of interest were cropped from the blotting images. The full-length blots are presented in Fig. S14(A).