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. 2024 Aug 7;632(8025):647–655. doi: 10.1038/s41586-024-07726-0

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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/.

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Extended Data Fig. 7 — (a) Cross-section view of tight junction formation phases on the cellular level. (1) ZO-1 (green) condensates nucleate at cell-cell contact sites. (2) Cells polarize and nucleated ZO-1 surface condensates grow around the apical interface (magenta). (3) Cell-cell contacts mature, and the tight junction belt closes and seals the tissue. (b) Mesoscale events during tight junction belt formation. (1) Nucleation of ZO-1 condensates leads to partitioning of junction proteins. (2) Interaction of nucleated condensates with the apical interface via PATJ induces condensate growth along the interface via a prewetting transition. Growing condensates fuse into a continuous tight junction belt. (3) Polymerization of tight junction strands establishes the tight trans-epithelial barrier. (c) Molecular interactions underlying prewetting of ZO-1 condensates along the apical interface. ZO-1 surface condensates nucleate by ZO-1 binding to receptors that bind in trans at cell-cell adhesion sites (JAMs, Claudins). PATJ binds to the apical membrane via the apical complex (Pals1). Preferential interactions of membrane bound ZO-1 and PATJ mediate enrichment of the ZO-1 – PATJ complex at the apical interface. Apical interface enrichment promotes growth of ZO-1 surface condensates via prewetting.