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. 2019 Dec 11;10:5650. doi: 10.1038/s41467-019-13538-y

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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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Fig. 3 — Flagellin administration alters the intestinal microbiota toward a lower pro-inflammatory state. a Fecal pro-inflammatory potential was analyzed using HEK 293 cells expressing mTLR5 or mTLR4 measuring bioactive flagellin and lipopolysaccharide, respectively. b Colonic myeloperoxidase quantification of 4-week old, wild-type C57BL/6 J mice after receiving either vehicle or 10 μg of flagellin by intraperitoneal injections weekly for 9 weeks. c–f Colonic microbiota localization analysis of wild type and μMT mice treated with PBS, Salmonella-derived flagellin, or Bacillus-derived flagellin. c, e Confocal microscopy analysis of colonic microbiota localization; Muc2 (green), actin (purple), bacteria (red), and DNA (blue). d, f Distances of closest bacteria to colonic intestinal epithelial cells (IEC) per condition over 2–3 high-powered fields per mouse. g Fecal bacterial load determined by qPCR analysis of 16 S bacterial DNA in the fecal contents of mice treated with PBS or flagellin. Data are the means ±S.E.M. Significance was determined using t test (*p ≤ 0.05 **p ≤ 0.01 ***p ≤ 0.001 ****p ≤ 0.0001, n.s. indicates non-significant). (N=4–5 mice from one out of three representative experiment). Source data are provided as a Source Data file.