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. 2023 Oct 5;14:6214. doi: 10.1038/s41467-023-41999-9

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© The Author(s) 2023

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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Fig. 4 — a Immunofluorescence staining and representative deconvoluted confocal images of control SKCO15 and CLDN23 overexpressing SKCO15 monolayers stained with anti-ZO1 (magenta & green) and either anti-CLDN3 (green), or anti-CLDN4 (magenta) antibodies. Scale bar: 20μm. Arrows point to TJ spike formation along cell-cell contacts. b Schematic representing the visualization of TJ strand formation employing super-resolution STED microscopy. Created with BioRender.com. c Left, representative super-resolution STED microscopy images in control SKCO15 IECs and CLDN23 overexpressing SKCO15 IEC monolayers stained with anti-ZO1 (magenta or green) and either anti-CLDN3 (green), or anti-CLDN4 (magenta) antibodies. Scale bar: 20 μm. Right, histograms showing cell–cell contact thickness. Results show the mean ± SD of two independent experiments. A total of 33 (CLDN3/ZO1) and 50 (CLDN4/ZO1) cell-cell contacts were analyzed for control cells, while 38 (CLDN3/ZO1) and 55 (CLDN4/ZO1) were analyzed for CLDN23 overexpressing cells. ****p < 0.0001; statistical analysis was done with two-tailed Student’s t test.