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. 2020 Jun 1;10:8903. doi: 10.1038/s41598-020-65867-4

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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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Infection by E. coli EV36-RFP of hCMEC cultures. (A) Representative fluorescent images showing E. coli EV36-RFP infection of hCMEC cultures at concentrations of 10⁶ CFU/ml (left) and 10⁷ CFU/ml (right). E. coli EV36-RFP fluorescence shown in red, DAPI in blue, and phalloidin in white (B) Flow cytometry histogram showing E. coli EV36-RFP infected hCMECs in red and non-infected hCMECs in grey, as detected by RFP fluorescence. (C,D) Mean percentages of E. coli EV36-RFP infected hCMECs (RFP-positive by flow cytometry) after a 1 h incubation period with concentrations of 10³ to 10⁷ CFU/ml (C), or 10⁷ CFU/ml sampled over time (D); + /− SD, n = 3 in each case.