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. 2019 Jun 19;9:8742. doi: 10.1038/s41598-019-45009-1

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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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Gut Leaching and Systemic Endotoxemia: Formalin-fixed, paraffin-embedded 5 μm intestinal slices from lean mouse control (No. of mice per group, N = 8), wild type mouse control (NAFLD) (N = 8), wild type mouse group exposed with Microcystin (CHOW + MC) (N = 8) and another NAFLD group of mice exposed with Microcystin (NAFLD + MC) (N = 8) were used for immunofluorescence imaging. (A) Immunofluorescence images depicting Claudin-2 (red) (i–iv) and (B) Occludin (red) (v–viii), counterstained with DAPI (blue) in LC, CHOW + MC, NAFLD and NAFLD + MC group of mice intestine. Images were taken at 20X magnification. (C) Morphometric analysis of Claudin-2 immunoreactivity. Y-axis shows % positive immunoreactive area (% ROI) (n = 5, analysis from five separate microscopic fields) (*p < 0.05) (D) Morphometric analysis of Occludin immunoreactivity. Y-axis shows % positive immunoreactive area (% ROI) (n = 5, analysis from five separate microscopic fields) (0.01 < *p < 0.05). (E) Systemic endotoxemia was measured by Chromogenic LAL assay in serum of LC (CHOW), CHOW + MC, NAFLD and NAFLD + MC group of mice.