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. 2024 Jun 7;25(7):3040–3063. doi: 10.1038/s44319-024-00170-1

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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/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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(A) H&E staining of representative kidney sections from wild-type and Pax3Cre-Dlg1^F/F mice. Black box denotes regions shown as magnified images on the right. Yellow arrows point to cystic tubules in the cortical region. (B–D) Immunofluorescence staining for cilia (acetylated α-tubulin, yellow) and quantification of ciliary length (C) and ciliary acetylated α-tubulin intensity (D) in kidney sections of wild-type (n = 3) and Pax3Cre-Dlg1^F/F (n = 4) mice at 4 months of age. Data are presented as mean ± SD and statistical analysis was performed using Student’s t-test. ****P < 0.0001. (E) Representative image of transwell filter-grown mCCD cell lines (mCh-DLG1: mCherry-DLG1). Cilia were visualized using acetylated α-tubulin antibody (AcTub, magenta), cell–cell contacts were visualized with E-cadherin antibody (green) and nuclei were stained with DAPI (blue). (F) Western blot analysis of total cell lysates of the indicated mCCD cells lines using antibodies against DLG1 and GAPDH (loading control). Molecular mass markers are shown in kDa to the left. (G, H) Quantification of ciliary length (G) and frequency (H) in the indicated transwell filter-grown mCCD lines. Ciliary length and ciliation rate were measured using a fully automated MATLAB script (see “Methods” for details). Graphs represent accumulated data from three independent experiments, and statistical analysis was performed using Mann–Whitney U test (unpaired, two-tailed). Error bars represent mean ± SD. **P < 0.01; ****P < 0.0001; ns, not statistically significant. Source data are available online for this figure.