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Loss of mammalian Abi1 causes myeloproliferative neoplasm; however, little is known about how the Abi family of actin-regulatory proteins regulates blood cell homeostasis. Here, we demonstrate that Drosophila Abi promotes plasmatocyte-to-crystal cell transdifferentiation but represses plasmatocyte-to-lamellocyte transdifferentiation through Notch signaling. Consistent with a previously demonstrated role of clathrin-mediated endocytosis (CME) in Notch signaling activation, we find that Abi promotes Notch-CME by recruiting WASp and the Notch receptor to nascent sites of CME. Finally, we demonstrate that CME and crystal cell formation are inhibited by Abelson (Abl)-mediated phosphorylation of Abi but require PTP61F, a phosphatase that reverses this phosphorylation. Our findings identify Abi as a critical integrator of actin remodeling and Notch-CME and reveal opposing roles of Abl and PTP61F in regulating Abi activity to maintain blood cell homeostasis.<\/jats:p>","DOI":"10.1083\/jcb.202505091","type":"journal-article","created":{"date-parts":[[2025,11,25]],"date-time":"2025-11-25T07:21:14Z","timestamp":1764055274000},"update-policy":"https:\/\/doi.org\/10.1083\/jcb.crossmarkpolicy","source":"Crossref","is-referenced-by-count":1,"title":["<i>Drosophila<\/i>\n                    Abi maintains blood cell homeostasis by promoting clathrin-mediated endocytosis of Notch"],"prefix":"10.1083","volume":"225","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1546-5866","authenticated-orcid":true,"given":"Hyun Gwan","family":"Park","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04h9pn542","id-type":"ROR","asserted-by":"publisher"}],"name":"Seoul National University 1 Department of Cell and Developmental Biology and Dental Research Institute, , Seoul,","place":["Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-4564-7420","authenticated-orcid":true,"given":"Seunghwan","family":"Song","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04h9pn542","id-type":"ROR","asserted-by":"publisher"}],"name":"Interdisciplinary Program in Neuroscience, Seoul National University 2 , Seoul,","place":["Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4425-6476","authenticated-orcid":true,"given":"Joohyung","family":"Kim","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04h9pn542","id-type":"ROR","asserted-by":"publisher"}],"name":"Seoul National University 1 Department of Cell and Developmental Biology and Dental Research Institute, , Seoul,","place":["Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6620-6269","authenticated-orcid":true,"given":"Seungbok","family":"Lee","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/04h9pn542","id-type":"ROR","asserted-by":"publisher"}],"name":"Seoul National University 1 Department of Cell and Developmental Biology and Dental Research Institute, , Seoul,","place":["Korea"]},{"id":[{"id":"https:\/\/ror.org\/04h9pn542","id-type":"ROR","asserted-by":"publisher"}],"name":"Interdisciplinary Program in Neuroscience, Seoul National University 2 , Seoul,","place":["Korea"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"291","published-online":{"date-parts":[[2025,12,26]]},"reference":[{"key":"2025123115271245900_bib1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1005746","article-title":"Transdifferentiation and proliferation in two distinct hemocyte lineages in Drosophila melanogaster larvae after wasp infection","volume":"12","author":"Anderl","year":"2016","journal-title":"Plos Pathog."},{"key":"2025123115271245900_bib2","doi-asserted-by":"publisher","first-page":"367","DOI":"10.1534\/genetics.118.300223","article-title":"Drosophila as a genetic model for hematopoiesis","volume":"211","author":"Banerjee","year":"2019","journal-title":"Genetics"},{"key":"2025123115271245900_bib3","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1016\/j.ydbio.2020.03.018","article-title":"Context-specific functions of Notch in Drosophila blood cell progenitors","volume":"462","author":"Blanco-Obregon","year":"2020","journal-title":"Dev. 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