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. 2020 May 4;10:7475. doi: 10.1038/s41598-020-64424-3

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© The Author(s) 2020

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) Biofilm formation of EAEC ST strains. Bacteria were incubated in 96-well plates for 48 h, stained with crystal violet and absorbance was quantified (OD₅₉₅). (B) Cytotoxicity of EAEC infection. Death of T84 cells was quantified by Trypan Blue staining after 8 h of infection. Non-infected cells (NI) were included as controls. (C) Secretion of interleukin-8 (IL-8) by T84 cells infected with EAEC. IL-8 levels in supernatants were quantified by ELISA after 24 h of incubation. Results represent the mean ± SE from three independent experiments in duplicate. Data from groups of ST31 and ST40 isolates were analysed using Student’s unpaired t-test. P* < 0.05, P*** < 0.001.