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Proteins of the PAR and Crumbs complexes are pivotal to epithelial polarization, but the mechanistic understanding of polarization is challenging to reach, largely because numerous potential interactions between these proteins and others have been found, without a clear hierarchy in importance. We identify the regionalized and segregated organization of members of the PAR and Crumbs complexes at epithelial apical junctions by imaging endogenous proteins using stimulated\u2010emission\u2010depletion microscopy on Caco-2 cells, and human and murine intestinal samples. Proteins organize in submicrometric clusters, with PAR3 overlapping with the tight junction (TJ) while PALS1-PATJ and aPKC-PAR6\u03b2 form segregated clusters that are apical of the TJ and present in an alternated pattern related to actin organization. CRB3A is also apical of the TJ and partially overlaps with other polarity proteins. Of the numerous potential interactions identified between polarity proteins, only PALS1-PATJ and aPKC-PAR6\u03b2 are spatially relevant in the junctional area of mature epithelial cells, simplifying our view of how polarity proteins could cooperate to drive and maintain cell polarity.<\/jats:p>","DOI":"10.7554\/elife.62087","type":"journal-article","created":{"date-parts":[[2022,11,4]],"date-time":"2022-11-04T14:15:13Z","timestamp":1667571313000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.62087","source":"Crossref","is-referenced-by-count":19,"title":["Super-resolution imaging uncovers the nanoscopic segregation of polarity proteins in epithelia"],"prefix":"10.7554","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8305-7322","authenticated-orcid":true,"given":"Pierre","family":"Mangeol","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/035xkbk20","id-type":"ROR","asserted-by":"publisher"}],"name":"Aix-Marseille University, CNRS, UMR7288, Developmental Biology Institute of Marseille (IBDM)","place":["Marseille, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dominique","family":"Massey-Harroche","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/035xkbk20","id-type":"ROR","asserted-by":"publisher"}],"name":"Aix-Marseille University, CNRS, UMR7288, Developmental Biology Institute of Marseille (IBDM)","place":["Marseille, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fabrice","family":"Richard","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/035xkbk20","id-type":"ROR","asserted-by":"publisher"}],"name":"Aix-Marseille University, CNRS, UMR7288, Developmental Biology Institute of Marseille (IBDM)","place":["Marseille, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jean-Paul","family":"Concordet","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/02vjkv261","id-type":"ROR","asserted-by":"publisher"}],"name":"Laboratoire Structure et Instabilit\u00e9 des G\u00e9nomes, Mus\u00e9um National d\u2019Histoire Naturelle (MNHN), Institut National de la Sant\u00e9 et de la Recherche M\u00e9dicale (INSERM), U1154, Centre National de la Recherche Scientifique (CNRS)","place":["Paris, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1066-7506","authenticated-orcid":true,"given":"Pierre-Fran\u00e7ois","family":"Lenne","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/035xkbk20","id-type":"ROR","asserted-by":"publisher"}],"name":"Aix-Marseille University, CNRS, UMR7288, Developmental Biology Institute of Marseille (IBDM)","place":["Marseille, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andr\u00e9","family":"Le Bivic","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/035xkbk20","id-type":"ROR","asserted-by":"publisher"}],"name":"Aix-Marseille University, CNRS, UMR7288, Developmental Biology Institute of Marseille (IBDM)","place":["Marseille, France"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"4374","published-online":{"date-parts":[[2022,11,7]]},"reference":[{"key":"bib1","doi-asserted-by":"publisher","first-page":"4293","DOI":"10.1242\/jcs.03170","article-title":"Par3 acts as a molecular organizer to define the apical domain of chick neuroepithelial cells","volume":"119","author":"Afonso","year":"2006","journal-title":"Journal of Cell Science"},{"key":"bib2","doi-asserted-by":"publisher","first-page":"347","DOI":"10.1095\/biolreprod.110.084400","article-title":"Cell polarity regulator PARD6B is essential for trophectoderm formation in the preimplantation mouse embryo","volume":"83","author":"Alarcon","year":"2010","journal-title":"Biology of Reproduction"},{"key":"bib3","doi-asserted-by":"publisher","first-page":"614","DOI":"10.1016\/j.bbamem.2007.08.029","article-title":"Polarity complex proteins","volume":"1778","author":"Ass\u00e9mat","year":"2008","journal-title":"Biochimica et Biophysica Acta"},{"key":"bib4","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1016\/j.semcdb.2017.10.018","article-title":"Role of the Crumbs proteins in ciliogenesis, cell migration and actin organization","volume":"81","author":"Bazelli\u00e8res","year":"2018","journal-title":"Seminars in Cell & Developmental Biology"},{"key":"bib5","doi-asserted-by":"publisher","DOI":"10.1186\/s12864-018-4715-9","article-title":"New genomic data and analyses challenge the traditional vision of animal epithelium evolution","volume":"19","author":"Belahbib","year":"2018","journal-title":"BMC Genomics"},{"key":"bib6","doi-asserted-by":"publisher","first-page":"833","DOI":"10.1016\/S0092-8674(00)80593-0","article-title":"Discs lost, a novel multi-PDZ domain protein","volume":"96","author":"Bhat","year":"1999","journal-title":"Establishes and Maintains Epithelial Polarity. 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