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. 2023 Jun 20;14:3652. doi: 10.1038/s41467-023-39398-1

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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 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. 5 — a Representative confocal z-stacks showing x/z-axis cross sections of tissue obtained from Tvp23b^+/− and Tvp23b^−/− littermates; tissue (blue), 1-µm beads (red). b, c Normalized distances and penetrability to beads from colonic tissue (n = 8 per genotype, **P = 0.0072). d, e Visualization and quantification of microbiota localization relative to colonic mucosal surface using fluorescent in situ hybridization. d Sections were hybridized to a probe that recognizes 16S rRNA of all bacteria (green) and counterstained with DAPI (blue). e Average distance of inner mucus layer quantified from 16S FISH to close epithelium in at least 20 sections per mouse (n = 4 per genotype, ***P = 0.0006). f Representative mucus staining with lectin wheat germ agglutinin (WGA) and counterstains for tissue and bacteria (Syto9, blue). g Mucosa-associated bacteria were quantified by qPCR of 16S rRNA gene copy number in distal colon (n = 4 per genotype, ***P = 0.0001). Data (c, e, g) are expressed as means ± s.d. and significance was determined by unpaired Student t-test.