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Widely recognised as a paradigm for research in cellular microbiology,\n                    <jats:italic>S. flexneri<\/jats:italic>\n                    has emerged as important to study mechanisms of cell-autonomous immunity, including septin cage entrapment. Here, we use high-content high-resolution microscopy to monitor the dynamic and heterogeneous\n                    <jats:italic>S. flexneri<\/jats:italic>\n                    infection process by assessing multiple host and bacterial parameters (DNA replication, protein translation, T3SS activity). In the case of infected host cells, we report a reduction in DNA and protein synthesis together with morphological changes that suggest\n                    <jats:italic>S. flexneri<\/jats:italic>\n                    can induce cell-cycle arrest. We developed an artificial intelligence image analysis approach using convolutional neural networks to reliably quantify, in an automated and unbiased manner, the recruitment of SEPT7 to intracellular bacteria. We discover that heterogeneous SEPT7 assemblies are recruited to bacteria with increased T3SS activation. Our automated microscopy workflow is useful to illuminate diverse host and bacterial interactions at the single-cell and population level, and to fully characterise the intracellular microenvironment controlling the\n                    <jats:italic>S. flexneri<\/jats:italic>\n                    infection process.\n                  <\/jats:p>","DOI":"10.7554\/elife.97495","type":"journal-article","created":{"date-parts":[[2024,6,3]],"date-time":"2024-06-03T20:25:14Z","timestamp":1717446314000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.97495","source":"Crossref","is-referenced-by-count":4,"title":["High-content high-resolution microscopy and deep learning-assisted analysis reveals host and bacterial heterogeneity during Shigella infection"],"prefix":"10.7554","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0289-738X","authenticated-orcid":true,"given":"Ana Teresa","family":"L\u00f3pez-Jim\u00e9nez","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00a0jsq62","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Infection Biology, London School of Hygiene and Tropical Medicine","place":["London, United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"},{"role":"corresponding-author","vocabulary":"crossref"},{"role":"conceptualization","vocabulary":"credit"},{"role":"data-curation","vocabulary":"credit"},{"role":"formal-analysis","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"project-administration","vocabulary":"credit"},{"role":"software","vocabulary":"credit"},{"role":"validation","vocabulary":"credit"},{"role":"visualization","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6304-3292","authenticated-orcid":true,"given":"Dominik","family":"Brokatzky","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00a0jsq62","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Infection Biology, London School of Hygiene and Tropical Medicine","place":["London, United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"},{"role":"conceptualization","vocabulary":"credit"},{"role":"investigation","vocabulary":"credit"},{"role":"methodology","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-7519-1841","authenticated-orcid":true,"given":"Kamla","family":"Pillay","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00a0jsq62","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Infection Biology, London School of Hygiene and Tropical Medicine","place":["London, United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"},{"role":"investigation","vocabulary":"credit"}]},{"given":"Tyrese","family":"Williams","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00a0jsq62","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Infection Biology, London School of Hygiene and Tropical Medicine","place":["London, United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"},{"role":"investigation","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9115-7120","authenticated-orcid":true,"given":"Gizem","family":"\u00d6zbaykal G\u00fcler","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00a0jsq62","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Infection Biology, London School of Hygiene and Tropical Medicine","place":["London, United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"},{"role":"software","vocabulary":"credit"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7286-6503","authenticated-orcid":true,"given":"Serge","family":"Mostowy","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00a0jsq62","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Infection Biology, London School of Hygiene and Tropical Medicine","place":["London, United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"},{"role":"corresponding-author","vocabulary":"crossref"},{"role":"conceptualization","vocabulary":"credit"},{"role":"funding-acquisition","vocabulary":"credit"},{"role":"supervision","vocabulary":"credit"},{"role":"writing-original-draft","vocabulary":"credit"},{"role":"writing-review-editing","vocabulary":"credit"}]}],"member":"4374","published-online":{"date-parts":[[2025,10,8]]},"reference":[{"key":"bib1","doi-asserted-by":"publisher","first-page":"497","DOI":"10.1038\/nrmicro3491","type":"journal-article","article-title":"A functional perspective on phenotypic heterogeneity in microorganisms","volume":"13","author":"Ackermann","year":"2015","journal-title":"Nature Reviews. 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