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. 2020 Aug 3;11(8):647. doi: 10.1038/s41419-020-02822-1

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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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Fig. 7 — a Upper left picture shows three IBs in a WT mouse. Arrow points to one adhesion scored as 6 (grade 2; tenacity 4). Lower left picture shows three adhesions (arrows) in a CAV1^−/− mouse scored as 6 (left; grade 2; tenacity 4), 7 (middle; grade 3; tenacity 4) and 9 (right; grade 4; tenacity 5), respectively. Upper right Haematoxylin&Eosin (H&E) image shows an IB without adhesion formation in a WT mouse. Lower right H&E image shows a parietal IB adhered to pancreas in a CAV1^−/− mouse. Quantification of adhesions shows significant differences between WT and CAV1^−/− mice. b Parietal tissues adjacent to IB areas were analyzed by double immunofluorescence for pan-cytokeratin (green) and FSP-1 (red). A representative picture of a WT mouse shows a mainly preserved MC monolayer in an IB adjacent area. Below, a representative image of a CAV1^−/− mouse shows double pan-cytokeratin/FSP-1 positive fibroblasts accumulated in the submesothelial zone of an adhesion. Nuclear 4′,6-diamidino-2-fenilindol (DAPI) is represented in blue. c Serial sections corresponding to IB areas were analyzed by immunohistochemistry for pan-cytokeratin and α-SMA. Representative pictures of a WT mouse show a preserved MC monolayer (pan-cytokeratin positive; α-SMA negative) in the adhesion zone. Serial tissue sections of a CAV1^−/− mouse show a severe adhesion area formed between parietal peritoneum and pancreatic tissue. Double pan-cytokeratin/α-SMA positive MCs are located into the submesothelial zone. Right graphics show submesothelial pan-cytokeratin and α-SMA staining quantifications. Submesothelial staining for pan-cytokeratin and α-SMA were significantly increased in CAV1^−/− compared to WT mice. Lower graphic shows a significant correlation between submesothelial pan-cytokeratin and α-SMA stains. d Peritoneal tissues were analyzed for phospho-Smad3 immunohistochemical staining, showing a significant increase of submesothelial positive areas in CAV1^−/− as compared to WT mice. e Similarly, YAP staining was significantly increased in submesothelial zones of CAV1^−/− as compared to WT mice. Graphic bars represent the mean ± SEM. Arrows point to the adhesion zone. IB ischemic button, PP parietal peritoneum, Adh adhesion, VP visceral peritoneum, P pancreas.