{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T04:02:39Z","timestamp":1784606559840,"version":"3.55.0"},"reference-count":29,"publisher":"Oxford University Press (OUP)","issue":"2","license":[{"start":{"date-parts":[[2026,4,25]],"date-time":"2026-04-25T00:00:00Z","timestamp":1777075200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["23\u00a0K08732"],"award-info":[{"award-number":["23\u00a0K08732"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["21\u00a0K06854"],"award-info":[{"award-number":["21\u00a0K06854"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["21\u00a0K09419"],"award-info":[{"award-number":["21\u00a0K09419"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,8,3]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The small GTPase RhoA has diverse cellular functions, including in actin fiber reorganization. However, its role in angiogenesis remains elusive. To reveal this, we generated mice with endothelium-specific constitutive or tamoxifen-inducible conditional knockout (cKO) of RhoA. Both constitutive and inducible RhoA cKO mice all died by embryonic day 11.5. They displayed severe abnormalities in cephalic plexus vasculature and intersomitic vessels. Inducible RhoA cKO mice also had abnormal lymphatic vessels at mid-stage embryogenesis and showed significant reductions in radial outgrowth and density of vascular plexus in the retina on postnatal day 10. Blood flow recovery and vascular repair in ischemic hindlimbs were also impaired in adult inducible RhoA cKO mice. The expression of a stalk cell marker Notch1 was reduced, but that of a tip cell marker Dll4 was unchanged in the branched vessels of RhoA cKO embryos. By luciferase analysis and a database search, KLF2 was identified as a negative regulator of Notch1 expression by binding to the NOTCH1 gene promoter. KLF2 was phosphorylated by the RhoA\u2013ROCK pathway, which inhibited KLF2 nuclear translocation, resulting in increases in Notch1 transcription and expression. Endothelial RhoA supports endothelial differentiation through KLF2\u2013Notch1 signaling and promotes embryonic and postnatal vascular development.<\/jats:p>","DOI":"10.1093\/jb\/mvag032","type":"journal-article","created":{"date-parts":[[2026,4,23]],"date-time":"2026-04-23T11:42:47Z","timestamp":1776944567000},"page":"121-133","source":"Crossref","is-referenced-by-count":0,"title":["Endothelial RhoA is essential for embryonic and postnatal vascular development by regulating KLF2\u2013Notch1 signaling"],"prefix":"10.1093","volume":"180","author":[{"given":"Wondwossen Wale","family":"Tesega","sequence":"first","affiliation":[{"name":"Division of Molecular Medical Biochemistry, Department of Biochemistry and Molecular Biology, Shiga University of Medical Science , Seta Tsukinowa-cho, Otsu, Shiga 520-2192 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Akio","family":"Shimizu","sequence":"additional","affiliation":[{"name":"Division of Molecular Medical Biochemistry, Department of Biochemistry and Molecular Biology, Shiga University of Medical Science , Seta Tsukinowa-cho, Otsu, Shiga 520-2192 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Taha","family":"Hussain","sequence":"additional","affiliation":[{"name":"Division of Molecular Medical Biochemistry, Department of Biochemistry and Molecular Biology, Shiga University of Medical Science , Seta Tsukinowa-cho, Otsu, Shiga 520-2192 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fumiya","family":"Sawasaki","sequence":"additional","affiliation":[{"name":"Division of Molecular Medical Biochemistry, Department of Biochemistry and Molecular Biology, Shiga University of Medical Science , Seta Tsukinowa-cho, Otsu, Shiga 520-2192 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lucia","family":"Sugawara","sequence":"additional","affiliation":[{"name":"Division of Molecular Medical Biochemistry, Department of Biochemistry and Molecular Biology, Shiga University of Medical Science , Seta Tsukinowa-cho, Otsu, Shiga 520-2192 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8662-4636","authenticated-orcid":false,"given":"Shoma","family":"Matsumoto","sequence":"additional","affiliation":[{"name":"Department of Stem Cells and Human Disease Models, Research Center for Animal Life Science, Shiga University of Medical Science , Seta Tsukinowa-cho, Otsu, Shiga 520-2192 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0645-6183","authenticated-orcid":false,"given":"Masatsugu","family":"Ema","sequence":"additional","affiliation":[{"name":"Department of Stem Cells and Human Disease Models, Research Center for Animal Life Science, Shiga University of Medical Science , Seta Tsukinowa-cho, Otsu, Shiga 520-2192 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Akira","family":"Sato","sequence":"additional","affiliation":[{"name":"Division of Molecular Medical Biochemistry, Department of Biochemistry and Molecular Biology, Shiga University of Medical Science , Seta Tsukinowa-cho, Otsu, Shiga 520-2192 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6750-1668","authenticated-orcid":false,"given":"Hisakazu","family":"Ogita","sequence":"additional","affiliation":[{"name":"Division of Molecular Medical Biochemistry, Department of Biochemistry and Molecular Biology, Shiga University of Medical Science , Seta Tsukinowa-cho, Otsu, Shiga 520-2192 ,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2026,4,25]]},"reference":[{"key":"2026072023172485700_ref1","doi-asserted-by":"crossref","first-page":"1172114","DOI":"10.3389\/fcell.2023.1172114","article-title":"Endothelial-derived angiocrine factors as instructors of embryonic development","volume":"11","author":"Bishop","year":"2023","journal-title":"Front Cell Dev Biol"},{"key":"2026072023172485700_ref2","doi-asserted-by":"crossref","first-page":"115","DOI":"10.3345\/cep.2021.00773","article-title":"Retinopathy of prematurity: a review of epidemiology and current treatment strategies","volume":"65","author":"Hong","year":"2022","journal-title":"Clin Exp Pediatr"},{"key":"2026072023172485700_ref3","doi-asserted-by":"crossref","first-page":"836","DOI":"10.1002\/1873-3468.14578","article-title":"RhoA regulation in space and time","volume":"597","author":"Seze","year":"2023","journal-title":"FEBS Lett."},{"key":"2026072023172485700_ref4","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1002\/cm.20472","article-title":"Rho-kinase\/ROCK: a key regulator of the cytoskeleton and cell polarity","volume":"67","author":"Amano","year":"2010","journal-title":"Cytoskeleton"},{"key":"2026072023172485700_ref5","doi-asserted-by":"crossref","first-page":"471","DOI":"10.1093\/cvr\/cvt075","article-title":"Localized RhoA GTPase activity regulates dynamics of endothelial monolayer integrity","volume":"99","author":"Szulcek","year":"2013","journal-title":"Cardiovasc. Res."},{"key":"2026072023172485700_ref6","doi-asserted-by":"crossref","first-page":"1874","DOI":"10.1073\/pnas.0308525100","article-title":"Rho activity critically and selectively regulates endothelial cell organization during angiogenesis","volume":"101","author":"Hoang","year":"2004","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2026072023172485700_ref7","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pone.0002853","article-title":"Three-dimensional analysis of vascular development in the mouse embryo","volume":"3","author":"Walls","year":"2008","journal-title":"PLoS One"},{"key":"2026072023172485700_ref8","doi-asserted-by":"crossref","DOI":"10.1101\/cshperspect.a006569","article-title":"VEGF and notch in tip and stalk cell selection","volume":"3","author":"Blanco","year":"2013","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"2026072023172485700_ref9","doi-asserted-by":"crossref","first-page":"894","DOI":"10.24976\/Discov.Med.202537196.79","article-title":"Puerarin inhibits the development of thyroid cancer through KLF2\/NOTCH1 signaling","volume":"37","author":"Zhou","year":"2025","journal-title":"Discov. Med."},{"key":"2026072023172485700_ref10","doi-asserted-by":"crossref","first-page":"803","DOI":"10.1038\/cr.2013.42","article-title":"FBW7 regulates endothelial functions by targeting KLF2 for ubiquitination and degradation","volume":"23","author":"Wang","year":"2013","journal-title":"Cell Res."},{"key":"2026072023172485700_ref11","doi-asserted-by":"crossref","first-page":"e17","DOI":"10.1161\/ATVBAHA.118.310223","article-title":"Vascular development","volume":"38","author":"Majesky","year":"2018","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"2026072023172485700_ref12","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1161\/01.ATV.0000054198.68894.88","article-title":"Involvement of RhoA\/rho kinase signaling in VEGF-induced endothelial cell migration and angiogenesis in vitro","volume":"23","author":"Nieuw Amerongen","year":"2003","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"2026072023172485700_ref13","doi-asserted-by":"crossref","first-page":"776","DOI":"10.1038\/nature05571","article-title":"Dll4 signalling through Notch1 regulates formation of tip cells during angiogenesis","volume":"445","author":"Hellstr\u00f6m","year":"2007","journal-title":"Nature"},{"key":"2026072023172485700_ref14","doi-asserted-by":"crossref","first-page":"4354","DOI":"10.1182\/blood-2005-08-3465","article-title":"KLF2 provokes a gene expression pattern that establishes functional quiescent differentiation of the endothelium","volume":"107","author":"Dekker","year":"2006","journal-title":"Blood"},{"key":"2026072023172485700_ref15","doi-asserted-by":"crossref","first-page":"845","DOI":"10.1016\/j.devcel.2006.09.006","article-title":"Klf2 is an essential regulator of vascular hemodynamic forces in vivo","volume":"11","author":"Lee","year":"2006","journal-title":"Dev. Cell"},{"key":"2026072023172485700_ref16","doi-asserted-by":"crossref","first-page":"2996","DOI":"10.1101\/gad.11.22.2996","article-title":"The LKLF transcription factor is required for normal tunica media formation and blood vessel stabilization during murine embryogenesis","volume":"11","author":"Kuo","year":"1997","journal-title":"Genes Dev."},{"key":"2026072023172485700_ref17","doi-asserted-by":"crossref","first-page":"20522","DOI":"10.1074\/jbc.M109.025346","article-title":"Kruppel-like factor 2 inhibits hypoxia-inducible factor 1alpha expression and function in the endothelium","volume":"284","author":"Kawanami","year":"2009","journal-title":"J. Biol. Chem."},{"key":"2026072023172485700_ref18","doi-asserted-by":"crossref","first-page":"2487","DOI":"10.1253\/circj.CJ-15-0283","article-title":"High concentrations of uric acid inhibit angiogenesis via regulation of the Kr\u00fcppel-like factor 2-vascular endothelial growth factor-A axis by miR-92a","volume":"79","author":"Yu","year":"2015","journal-title":"Circ. J."},{"key":"2026072023172485700_ref19","doi-asserted-by":"crossref","first-page":"11666","DOI":"10.1038\/s41598-019-48053-z","article-title":"Endothelial RhoA GTPase is essential for in vitro endothelial functions but dispensable for physiological in vivo angiogenesis","volume":"9","author":"Zahra","year":"2019","journal-title":"Sci. Rep."},{"key":"2026072023172485700_ref20","first-page":"3058","article-title":"Cdc42 is required for cytoskeletal support of endothelial cell adhesion during blood vessel formation in mice","volume":"142","author":"Barry","year":"2015","journal-title":"Development"},{"key":"2026072023172485700_ref21","doi-asserted-by":"crossref","first-page":"1829","DOI":"10.1096\/fj.07-096438","article-title":"An essential role for Rac1 in endothelial cell function and vascular development","volume":"22","author":"Tan","year":"2008","journal-title":"FASEB J."},{"key":"2026072023172485700_ref22","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1007\/s10456-022-09831-y","article-title":"Angiopoietin-2-induced lymphatic endothelial cell migration drives lymphangiogenesis via the \u03b21 integrin-RhoA-formin axis","volume":"25","author":"Akwii","year":"2022","journal-title":"Angiogenesis"},{"key":"2026072023172485700_ref23","doi-asserted-by":"crossref","first-page":"1426","DOI":"10.1039\/c3ib40085a","article-title":"The mouse retina in 3D: quantification of vascular growth and remodeling","volume":"5","author":"Milde","year":"2013","journal-title":"Integr. Biol."},{"key":"2026072023172485700_ref24","doi-asserted-by":"crossref","first-page":"726","DOI":"10.1038\/s41467-017-00738-7","article-title":"Transcriptional regulation of endothelial cell behavior during sprouting angiogenesis","volume":"8","author":"Jeong","year":"2017","journal-title":"Nat. Commun."},{"key":"2026072023172485700_ref25","doi-asserted-by":"crossref","first-page":"679","DOI":"10.1242\/jcs.064964","article-title":"Rho GTPases and their role in organizing the actin cytoskeleton","volume":"124","author":"Sit","year":"2011","journal-title":"J. Cell Sci."},{"key":"2026072023172485700_ref26","doi-asserted-by":"crossref","first-page":"1473","DOI":"10.1182\/blood-2014-12-616169","article-title":"Cytokinesis failure in RhoA-deficient mouse erythroblasts involves actomyosin and midbody dysregulation and triggers p53 activation","volume":"126","author":"Konstantinidis","year":"2015","journal-title":"Blood"},{"key":"2026072023172485700_ref27","doi-asserted-by":"crossref","first-page":"911","DOI":"10.1016\/j.neuron.2011.12.030","article-title":"A radial glia-specific role of RhoA in double cortex formation","volume":"73","author":"Cappello","year":"2012","journal-title":"Neuron"},{"key":"2026072023172485700_ref28","doi-asserted-by":"crossref","first-page":"1071","DOI":"10.1038\/s42003-022-04042-z","article-title":"Vascular smooth muscle RhoA counteracts abdominal aortic aneurysm formation by modulating MAP 4K4 activity","volume":"5","author":"Molla","year":"2022","journal-title":"Commun Biol"},{"key":"2026072023172485700_ref29","doi-asserted-by":"crossref","DOI":"10.1016\/j.jbc.2023.102993","article-title":"RhoA rescues cardiac senescence by regulating Parkin-mediated mitophagy","volume":"299","author":"Soh","year":"2023","journal-title":"J. Biol. Chem."}],"container-title":["The Journal of Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/academic.oup.com\/jb\/advance-article-pdf\/doi\/10.1093\/jb\/mvag032\/68156480\/mvag032.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jb\/article-pdf\/180\/2\/121\/68156480\/mvag032.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/academic.oup.com\/jb\/article-pdf\/180\/2\/121\/68156480\/mvag032.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T03:17:40Z","timestamp":1784603860000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/jb\/article\/180\/2\/121\/8662153"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,25]]},"references-count":29,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2026,4,25]]},"published-print":{"date-parts":[[2026,8,3]]}},"URL":"https:\/\/doi.org\/10.1093\/jb\/mvag032","relation":{},"ISSN":["0021-924X","1756-2651"],"issn-type":[{"value":"0021-924X","type":"print"},{"value":"1756-2651","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2026,8]]},"published":{"date-parts":[[2026,4,25]]}}}