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. 2018 Feb 27;8:3719. doi: 10.1038/s41598-018-22017-1

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© The Author(s) 2018

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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EspF and Map inhibit de novo tight junction assembly. (A–D) Untransfected, EGFP, EGFP-Tir, EGFP-EspF or EGFP-Map cells were grown on permeable filters in low calcium (<5 µM) medium for 18–20 hours and then switched to normal calcium (~1.8 mM) medium to initiate junction assembly. TER was measured hourly for 24 hours. EGFP-1, EGFP-2; Tir-1, Tir-2; EspF-1, EspF-2, Map-1 and Map-2 represent biological replicates. Data are represented as means ± s.e.m. from three independent experiments with 4 filters (n = 4) taken for each cell line; **indicates p-value < 0.001. (E,F) The paracellular flux of 4 kDa and 70 kDa dextran tracers was measured in untransfected, EGFP, EGFP-Tir, EGFP-EspF and EGFP-Map cell lines. Shown are fold changes calculated with respect to untransfected cells (normalized to 1). Error bars represent means ± s.e.m from three independent experiments; *represents p-value < 0.01 and **represents p-value < 0.001.