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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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Claudin proteins are altered in response to LPS in Caco-2 cells. Treatment of Caco-2 cells with LPS for 7 days induces changes in claudin gene and protein expression. (a) qRT-PCR showed experimental NEC decreased gene expression of claudins 1, 3, and 4 (n = 3/5/5, respectively), and increased claudin 2 expression (n = 9). Genes were normalized to GAPDH. (b) Western blot analysis showing ratio of claudin protein expression in experimental NEC cells to same-day controls at days 0 and 7. Protein expression of claudin 1 and 3 is decreased in experimental NEC, while claudin 2 and 4 expression is increased. Below, representative immunoblots (full blots included in Supplementary Fig. S1) probed with antibodies against each claudin on day 0 and 7 in control (C) and LPS-treated cells (N); blots were reprobed with antibodies against β-Actin; analyzed in at least three separate experiments. All values are mean ± SEM (*p < 0.05).