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. 2019 Mar 21;9:4982. doi: 10.1038/s41598-019-41442-4

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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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Increased permeability in experimental NEC. (a) Caco-2 cells treated with LPS for 7 days (NEC) have increased permeability to 3 kDa FITC-dextran (n = 15). (b) LPS treatment in Caco-2 cells decreased TEER progressively from day 1 (−1% control vs. −14% NEC, ***p < 0.001) through day 7 (−13% vs. −30%, **p < 0.01) (n = 23). (c) Caco-2 cell immunofluorescent DNA fragmentation staining apoptotic cell count per 1000 cells showed no difference between controls and LPS-treated cells (p = 0.32, n = 10). (d) The combination of hypoxia and formula feedings without (control) or with Cronobacter sakasakii bacteria (NEC) results in increased translocation of gavage fed FITC-dextran through the intestinal epithelium into the blood. Serum from rat pups was collected and permeability was assessed by FITC-assay (n = 9 control, 6 NEC). Outliers greater than 2 standard deviations were excluded. All values are mean ± SEM (*p < 0.05, **p < 0.01, ***p < 0.001).