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. 2024 Feb 27;55:25. doi: 10.1186/s13567-024-01266-1

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Illustration of the experimental setup. Equine 3D enteroids and 2D monolayers were generated from tissues originating from two individual horses. A Equine 3D enteroids from horse 1 cultured in plain growth medium or stimulated with eqIL-4 and IL-13 were used to optimize the labelling conditions for immunofluorescence microscopy (IF). B Equine 3D enteroids from horse 1 and 2 were disrupted to single cells, cultured as 2D monolayers on transwell supports and monitored by TEER. The monolayers were grown in plain growth medium or in the presence of eqIL-4/IL-13 before exposure to different combinations of P. univalens, cyathostomin or S. vulgaris larvae. Gene expression of cytokines/chemokines and cell lineage markers were examined by qPCR analysis, and verified by IF and SEM. C To enable live-cell imaging during exposure to nematode larvae, enteroid monolayers originating from horse 1 and 2 were cultured in AICs built to optimize the optical conditions for DIC microscopy of the apical epithelial surface. These AIC-grown monolayers were exposed to P. univalens, cyathostomin and S. vulgaris larvae and compared to parallel controls. Illustration created with https://www.BioRender.com.