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. 2017 Jun 7;7:3007. doi: 10.1038/s41598-017-03020-4

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© The Author(s) 2017

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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M. marinum ΔwhiB4 was less potent inducing TNF. RAW 264.7 macrophage cells were infected with mycobacteria at MOI of 10. At 3, 12, and 24-hr post infection, cell-free supernatants were collected and the levels of TNF were determined by enzyme-linked immunosorbent assay (ELISA). Data are plotted as mean ± SEM (n = 3), and are representative of two independent experiments. The level of TNF secretion induced by ΔwhiB4 was significantly lower than that of WT or the complemented strain (***p < 0.001; two-way ANOVA).