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. 2022 Jun 30;13:3780. doi: 10.1038/s41467-022-31587-8

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© The Author(s) 2022

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. 9 — a The dynamin inhibitor MiTMAB (20 μM) prevents EGFP-claudin-2 (cyan) removal from tight junctions following induction (+Dox) of mCherry-claudin-4 (magenta) expression. Maximum projection images representative of 5 independent experiments. Scale: 20 µm, 5 µm, 2 µm. b Dynamin inhibition (dark symbols) reduces numbers of EGFP-claudin-2 vesicles (−Dox, cyan) in the absence of claudin-4 and blocks EGFP-claudin-2 endocytosis triggered by mCherry-claudin-4 expression (+Dox, magenta symbols). n = 8. Each data point represents 4–5 cells, representative of 3 independent experiments. 1-way ANOVA. ***P < 0.0001. c MiTMAB (dark symbols) increases EGFP-claudin-2 expression at the tight junction in the absence of claudin-4 (−Dox, cyan symbols) and prevents EGFP-claudin-2 endocytosis after mCherry-claudin-4 expression (+Dox, magenta symbols). n = 9. Each data point represents 4–5 cells, representative of 5 independent experiments. 1-way ANOVA. ***P < 0.0001. Scale: 5 µm. d Endocytic inhibitors (dark symbols) MiTMAB, chlorpromazine (CPZ, 30 μM), or dynamin inhibitory peptide (DIP, 25 μM) do not affect TER in the absence of claudin-4 (cyan symbols) but fail to prevent TER increases induced by mCherry-claudin-4 expression (+Dox, magenta symbols). n = 3, representative of 3 independent experiments. Two-tail unpaired t-test. ***P < 0.0001. e MitMAB (dark symbols) does not affect EGFP-claudin-2 anchoring in the absence of claudin-4 (+Dox, magenta symbols) but is insufficient to prevent EGFP-claudin-2 mobilization after mCherry-claudin-4 expression (+Dox, magenta symbols). n = 4, representative of 3 independent experiments. 1-way ANOVA. **P = 0.0063, ***P < 0.001. Data are presented as mean ± SD and included in the Source Data file.