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. 2021 Mar 15;11:5945. doi: 10.1038/s41598-021-85399-9

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© The Author(s) 2021

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/.

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Dual immunofluorescence staining for CARMIL2 protein and CK18 on sigmoid FFPE sections of healthy control, IBD control, and three biallelic CARMIL2 variant carriers diagnosed with IBD. DAPI counterstaining was used to visualize nuclei. (A) Composite image, where green staining indicates CARMIL2, red represents cytokeratin 18 (CK18), a marker for single layer epithelial cells, and blue marks nuclear DAPI stain. In Patient 1 and Patient 3, carriers of the two protein-truncating variants (respectively p.C155VfsTer54 and p.E978*), CARMIL2 signal was almost absent in sigmoid sections. In Patient 4, carrier of missense variant p.D623E, immunofluorescence staining was weaker than the controls particularly in the mucosal layer. (B) Single-label immunofluorescence for CK18 (red). (C) Single-label immunofluorescence for CARMIL2 (green).