{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,14]],"date-time":"2026-02-14T05:12:27Z","timestamp":1771045947182,"version":"3.50.1"},"reference-count":87,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2024,2,5]],"date-time":"2024-02-05T00:00:00Z","timestamp":1707091200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Leonard H. Berenfield Graduate Fellowship"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Endothelial cells (ECs) respond to concurrent stimulation by biochemical factors and wall shear stress (SS) exerted by blood flow. Disruptions in flow-induced responses can result in remodeling issues and cardiovascular diseases, but the detailed mechanisms linking flow-mechanical cues and biochemical signaling remain unclear. Activin receptor-like kinase 1 (ALK1) integrates SS and ALK1-ligand cues in ECs; ALK1 mutations cause hereditary hemorrhagic telangiectasia (HHT), marked by arteriovenous malformation (AVM) development. However, the mechanistic underpinnings of ALK1 signaling modulation by fluid flow and the link to AVMs remain uncertain. We recorded EC responses under varying SS magnitudes and ALK1 ligand concentrations by assaying pSMAD1\/5\/9 nuclear localization using a custom multi-SS microfluidic device and a custom image analysis pipeline. We extended the previously reported synergy between SS and BMP9 to include BMP10 and BMP9\/10. Moreover, we demonstrated that this synergy is effective even at extremely low SS magnitudes (0.4 dyn\/cm2) and ALK1 ligand range (femtogram\/mL). The synergistic response to ALK1 ligands and SS requires the kinase activity of ALK1. Moreover, ALK1\u2019s basal activity and response to minimal ligand levels depend on endocytosis, distinct from cell\u2013cell junctions, cytoskeleton-mediated mechanosensing, or cholesterol-enriched microdomains. However, an in-depth analysis of ALK1 receptor trafficking\u2019s molecular mechanisms requires further investigation.<\/jats:p>","DOI":"10.3390\/cells13030285","type":"journal-article","created":{"date-parts":[[2024,2,5]],"date-time":"2024-02-05T08:41:43Z","timestamp":1707122503000},"page":"285","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Shear Stress and Sub-Femtomolar Levels of Ligand Synergize to Activate ALK1 Signaling in Endothelial Cells"],"prefix":"10.3390","volume":"13","author":[{"given":"Ya-Wen","family":"Cheng","sequence":"first","affiliation":[{"name":"Department of Bioengineering, Swanson School of Engineering, University of Pittsburgh, Pittsburgh, PA 15261, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anthony R.","family":"Anzell","sequence":"additional","affiliation":[{"name":"Department of Human Genetics, School of Public Health, University of Pittsburgh, Pittsburgh, PA 15261, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stefanie A.","family":"Morosky","sequence":"additional","affiliation":[{"name":"Department of Human Genetics, School of Public Health, University of Pittsburgh, Pittsburgh, PA 15261, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tristin A.","family":"Schwartze","sequence":"additional","affiliation":[{"name":"Department of Structural Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cynthia S.","family":"Hinck","sequence":"additional","affiliation":[{"name":"Department of Structural Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3320-8054","authenticated-orcid":false,"given":"Andrew P.","family":"Hinck","sequence":"additional","affiliation":[{"name":"Department of Structural Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1250-1705","authenticated-orcid":false,"given":"Beth L.","family":"Roman","sequence":"additional","affiliation":[{"name":"Department of Human Genetics, School of Public Health, University of Pittsburgh, Pittsburgh, PA 15261, USA"},{"name":"Heart, Lung, Blood and Vascular Medicine Institute, University of Pittsburgh, Pittsburgh, PA 15261, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2956-0437","authenticated-orcid":false,"given":"Lance A.","family":"Davidson","sequence":"additional","affiliation":[{"name":"Department of Bioengineering, Swanson School of Engineering, University of Pittsburgh, Pittsburgh, PA 15261, USA"},{"name":"Department of Developmental Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, USA"},{"name":"Department of Computational and Systems Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,2,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1152\/physrev.1995.75.3.519","article-title":"Flow-mediated endothelial mechanotransduction","volume":"75","author":"Davies","year":"1995","journal-title":"Physiol. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"H1209","DOI":"10.1152\/ajpheart.01047.2006","article-title":"Mechanotransduction and endothelial cell homeostasis: The wisdom of the cell","volume":"292","author":"Chien","year":"2007","journal-title":"Am. J. Physiol. Heart Circ. Physiol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1983","DOI":"10.1253\/circj.CJ-09-0583","article-title":"Vascular mechanobiology: Endothelial cell responses to fluid shear stress","volume":"73","author":"Ando","year":"2009","journal-title":"Circ. J."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1126\/science.3941904","article-title":"Reductions in arterial diameter produced by chronic decreases in blood flow are endothelium-dependent","volume":"231","author":"Langille","year":"1986","journal-title":"Science"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1007\/s00441-008-0706-5","article-title":"Arterial versus venous endothelial cells","volume":"335","year":"2009","journal-title":"Cell Tissue Res."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1431","DOI":"10.1056\/NEJM199405193302008","article-title":"The emerging concept of vascular remodeling","volume":"330","author":"Gibbons","year":"1994","journal-title":"N. Engl. J. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1517\/14712598.1.2.253","article-title":"General aspects of anti-angiogenesis and cancer therapy","volume":"1","author":"Zogakis","year":"2001","journal-title":"Expert Opin. Biol. Ther."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"932","DOI":"10.1038\/nature04478","article-title":"Angiogenesis in life, disease and medicine","volume":"438","author":"Carmeliet","year":"2005","journal-title":"Nature"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1016\/j.bcp.2018.10.023","article-title":"Fluid shear stress sensing in vascular homeostasis and remodeling: Towards the development of innovative pharmacological approaches to treat vascular dysfunction","volume":"158","author":"Baeyens","year":"2018","journal-title":"Biochem. Pharmacol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1042\/BJ20110301","article-title":"Signal transduction by vascular endothelial growth factor receptors","volume":"437","author":"Koch","year":"2011","journal-title":"Biochem. J."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1038\/nature03952","article-title":"A mechanosensory complex that mediates the endothelial cell response to fluid shear stress","volume":"437","author":"Tzima","year":"2005","journal-title":"Nature"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1161\/01.RES.0000089257.94002.96","article-title":"Ligand-independent activation of vascular endothelial growth factor receptor 2 by fluid shear stress regulates activation of endothelial nitric oxide synthase","volume":"93","author":"Jin","year":"2003","journal-title":"Circ. Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"623769","DOI":"10.3389\/fphys.2020.623769","article-title":"Endothelial Cell Orientation and Polarity Are Controlled by Shear Stress and VEGF through Distinct Signaling Pathways","volume":"11","author":"Vion","year":"2020","journal-title":"Front. Physiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"975","DOI":"10.1083\/jcb.201408103","article-title":"Intramembrane binding of VE-cadherin to VEGFR2 and VEGFR3 assembles the endothelial mechanosensory complex","volume":"208","author":"Coon","year":"2015","journal-title":"J. Cell Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e04645","DOI":"10.7554\/eLife.04645","article-title":"Vascular remodeling is governed by a VEGFR3-dependent fluid shear stress set point","volume":"4","author":"Baeyens","year":"2015","journal-title":"eLife"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"e168352","DOI":"10.1172\/JCI168352","article-title":"SMAD4 maintains the fluid shear stress set point to protect against arterial-venous malformations","volume":"133","author":"Banerjee","year":"2023","journal-title":"J. Clin. Investig."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Alt, A., Miguel-Romero, L., Donderis, J., Aristorena, M., Blanco, F.J., Round, A., Rubio, V., Bernabeu, C., and Marina, A. (2012). Structural and functional insights into endoglin ligand recognition and binding. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0029948"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1158\/1535-7163.MCT-09-0650","article-title":"ALK1-Fc inhibits multiple mediators of angiogenesis and suppresses tumor growth","volume":"9","author":"Mitchell","year":"2010","journal-title":"Mol. Cancer Ther."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"27313","DOI":"10.1074\/jbc.M112.377960","article-title":"Specificity and structure of a high affinity activin receptor-like kinase 1 (ALK1) signaling complex","volume":"287","author":"Townson","year":"2012","journal-title":"J. Biol. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"6328","DOI":"10.1021\/bi300942x","article-title":"Structure of the Alk1 extracellular domain and characterization of its bone morphogenetic protein (BMP) binding properties","volume":"51","author":"Mahlawat","year":"2012","journal-title":"Biochemistry"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"813","DOI":"10.1016\/j.molcel.2004.07.016","article-title":"Structural basis of heteromeric smad protein assembly in TGF-beta signaling","volume":"15","author":"Chacko","year":"2004","journal-title":"Mol. Cell"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Hinck, A.P., Mueller, T.D., and Springer, T.A. (2016). Structural Biology and Evolution of the TGF-beta Family. Cold Spring Harb. Perspect. Biol., 8.","DOI":"10.1101\/cshperspect.a022103"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"805","DOI":"10.1161\/CIRCULATIONAHA.120.053047","article-title":"Defective flow-migration coupling causes arteriovenous malformations in hereditary hemorrhagic telangiectasia","volume":"144","author":"Park","year":"2021","journal-title":"Circulation"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1573","DOI":"10.1242\/dev.060467","article-title":"Interaction between alk1 and blood flow in the development of arteriovenous malformations","volume":"138","author":"Corti","year":"2011","journal-title":"Development"},{"key":"ref_25","first-page":"2593","article-title":"Alk1 controls arterial endothelial cell migration in lumenized vessels","volume":"143","author":"Rochon","year":"2016","journal-title":"Development"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"807","DOI":"10.1083\/jcb.201603106","article-title":"Defective fluid shear stress mechanotransduction mediates hereditary hemorrhagic telangiectasia","volume":"214","author":"Baeyens","year":"2016","journal-title":"J. Cell Biol."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Schimmel, K., Ali, M.K., Tan, S.Y., Teng, J., Do, H.M., Steinberg, G.K., Stevenson, D.A., and Spiekerkoetter, E. (2021). Arteriovenous malformations\u2014Current understanding of the pathogenesis with implications for treatment. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22169037"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"4539","DOI":"10.1007\/s00018-017-2636-4","article-title":"ALK1 signaling in development and disease: New paradigms","volume":"74","author":"Roman","year":"2017","journal-title":"Cell. Mol. Life Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1093\/cvr\/cvr232","article-title":"Pulmonary hypertension in adult Alk1 heterozygous mice due to oxidative stress","volume":"92","author":"Jerkic","year":"2011","journal-title":"Cardiovasc. Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"851","DOI":"10.1164\/rccm.200908-1284OC","article-title":"Clinical outcomes of pulmonary arterial hypertension in patients carrying an ACVRL1 (ALK1) mutation","volume":"181","author":"Girerd","year":"2010","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"333","DOI":"10.1002\/ajmg.a.33206","article-title":"Overlapping spectra of SMAD4 mutations in juvenile polyposis (JP) and JP-HHT syndrome","volume":"152A","author":"Gallione","year":"2010","journal-title":"Am. J. Med. Genet. A"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1007","DOI":"10.1242\/bio.20134622","article-title":"The nucleus of endothelial cell as a sensor of blood flow direction","volume":"2","author":"Tkachenko","year":"2013","journal-title":"Biol. Open"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"869","DOI":"10.1016\/j.jvs.2005.01.020","article-title":"Differential effects of orbital and laminar shear stress on endothelial cells","volume":"41","author":"Dardik","year":"2005","journal-title":"J. Vasc. Surg."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"3241","DOI":"10.1242\/dev.150904","article-title":"Endothelial cells respond to the direction of mechanical stimuli through SMAD signaling to regulate coronary artery size","volume":"144","author":"Poduri","year":"2017","journal-title":"Development"},{"key":"ref_35","first-page":"201","article-title":"Small but perfectly formed? Successes, challenges, and opportunities for microfluidics in the chemical and biological sciences","volume":"2","author":"Chiu","year":"2017","journal-title":"Chemnanomat"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3563","DOI":"10.1002\/elps.200305584","article-title":"Microfluidic devices fabricated in poly(dimethylsiloxane) for biological studies","volume":"24","author":"Sia","year":"2003","journal-title":"Electrophoresis"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"4373","DOI":"10.1039\/D0LC00738B","article-title":"Endothelial cell polarization and orientation to flow in a novel microfluidic multimodal shear stress generator","volume":"20","author":"Sonmez","year":"2020","journal-title":"Lab. Chip"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1177\/153857440303700107","article-title":"Arterial wall shear stress: Observations from the bench to the bedside","volume":"37","author":"Paszkowiak","year":"2003","journal-title":"Vasc. Endovasc. Surg."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"821","DOI":"10.1172\/JCI83083","article-title":"Endothelial fluid shear stress sensing in vascular health and disease","volume":"126","author":"Baeyens","year":"2016","journal-title":"J. Clin. Investig."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1161\/01.RES.0000141145.78900.44","article-title":"Influence of mechanical, cellular, and molecular factors on collateral artery growth (arteriogenesis)","volume":"95","author":"Heil","year":"2004","journal-title":"Circ. Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1083\/jcb.200210135","article-title":"Shear stress-induced endothelial cell polarization is mediated by Rho and Rac but not Cdc42 or PI 3-kinases","volume":"161","author":"Ridley","year":"2003","journal-title":"J. Cell Biol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"536","DOI":"10.1161\/01.RES.76.4.536","article-title":"Nitric oxide synthesis by cultured endothelial cells is modulated by flow conditions","volume":"76","author":"Noris","year":"1995","journal-title":"Circ. Res."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"2130","DOI":"10.1161\/ATVBAHA.113.301826","article-title":"Endothelial cell sensing of flow direction","volume":"33","author":"Wang","year":"2013","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Mendez, P.L., Obendorf, L., Jatzlau, J., Burdzinski, W., Reichenbach, M., Nageswaran, V., Haghikia, A., Stangl, V., Hiepen, C., and Knaus, P. (2022). Atheroprone fluid shear stress-regulated ALK1-Endoglin-SMAD signaling originates from early endosomes. BMC Biol., 20.","DOI":"10.1186\/s12915-022-01396-y"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of relative gene expression data using real-time quantitative PCR and the 2\u2212\u0394\u0394CT method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"9984","DOI":"10.1074\/jbc.274.15.9984","article-title":"Assignment of transforming growth factor beta1 and beta3 and a third new ligand to the type I receptor ALK-1","volume":"274","author":"Lux","year":"1999","journal-title":"J. Biol. Chem."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"1427","DOI":"10.1016\/j.str.2019.06.010","article-title":"Structural Adaptation in Its Orphan Domain Engenders Betaglycan with an Alternate Mode of Growth Factor Binding Relative to Endoglin","volume":"27","author":"Kim","year":"2019","journal-title":"Structure"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: An open-source platform for biological-image analysis","volume":"9","author":"Schindelin","year":"2012","journal-title":"Nat. Methods"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2661","DOI":"10.1021\/acsbiomaterials.1c00251","article-title":"Enhancement of endothelialization by topographical features is mediated by PTP1B-dependent endothelial adherens junctions remodeling","volume":"7","author":"Gorji","year":"2021","journal-title":"ACS Biomater. Sci. Eng."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"3545","DOI":"10.1038\/s41467-018-05851-9","article-title":"Junction-based lamellipodia drive endothelial cell rearrangements in vivo via a VE-cadherin-F-actin based oscillatory cell-cell interaction","volume":"9","author":"Paatero","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.ddmec.2007.06.001","article-title":"Membrane rafts in endothelial cells and their modulation by proatherogenic stimuli","volume":"4","author":"Patschan","year":"2007","journal-title":"Drug Discov. Today Dis. Mech."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1628","DOI":"10.1111\/j.1476-5381.2010.00993.x","article-title":"Emodin suppresses lipopolysaccharide-induced pro-inflammatory responses and NF-kappaB activation by disrupting lipid rafts in CD14-negative endothelial cells","volume":"161","author":"Meng","year":"2010","journal-title":"Br. J. Pharmacol."},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Motulsky, H., and Christopoulos, A. (2004). Fitting Models to Biological Data Using Linear and Nonlinear Regression: A Practical Guide to Curve Fitting, Oxford University Press.","DOI":"10.1093\/oso\/9780195171792.001.0001"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1002\/dvdy.395","article-title":"BMP9 and BMP10: Two close vascular quiescence partners that stand out","volume":"251","author":"Tillet","year":"2022","journal-title":"Dev. Dyn."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"373","DOI":"10.1089\/ars.2015.6493","article-title":"Endothelial mechanosignaling: Does one sensor fit all?","volume":"25","author":"Givens","year":"2016","journal-title":"Antioxid. Redox Signal."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"H1561","DOI":"10.1152\/ajpheart.00081.2004","article-title":"Measuring hemodynamic changes during mammalian development","volume":"287","author":"Jones","year":"2004","journal-title":"Am. J. Physiol. Heart Circ. Physiol."},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Olsen, O.E., Skj\u00e6rvik, A., St\u00f8rdal, B.F., Sundan, A., and Holien, T. (2017). TGF-\u03b2 contamination of purified recombinant GDF15. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0187349"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"e87","DOI":"10.1161\/ATVBAHA.119.313122","article-title":"Impaired SMAD1\/5 Mechanotransduction and Cx37 (Connexin37) expression enable pathological vessel enlargement and shunting","volume":"40","author":"Peacock","year":"2020","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_59","doi-asserted-by":"crossref","unstructured":"W\u00f6ltje, K., Jabs, M., and Fischer, A. (2015). Serum induces transcription of Hey1 and Hey2 genes by Alk1 but not Notch signaling in endothelial cells. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0120547"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"4388","DOI":"10.1016\/j.bmcl.2008.06.052","article-title":"Structure-activity relationship study of bone morphogenetic protein (BMP) signaling inhibitors","volume":"18","author":"Cuny","year":"2008","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Sanvitale, C.E., Kerr, G., Chaikuad, A., Ramel, M.-C., Mohedas, A.H., Reichert, S., Wang, Y., Triffitt, J.T., Cuny, G.D., and Yu, P.B. (2013). A new class of small molecule inhibitor of BMP signaling. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0062721"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"1024","DOI":"10.1016\/j.cub.2013.04.049","article-title":"Fluid shear stress on endothelial cells modulates mechanical tension across VE-cadherin and PECAM-1","volume":"23","author":"Conway","year":"2013","journal-title":"Curr. Biol."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1194\/jlr.TR119000383","article-title":"The ins and outs of lipid rafts: Functions in intracellular cholesterol homeostasis, microparticles, and cell membranes: Thematic Review Series: Biology of Lipid Rafts","volume":"61","author":"Ouweneel","year":"2020","journal-title":"J. Lipid Res."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"1311","DOI":"10.1016\/j.bbamem.2007.03.026","article-title":"Use of cyclodextrins to manipulate plasma membrane cholesterol content: Evidence, misconceptions and control strategies","volume":"1768","author":"Zidovetzki","year":"2007","journal-title":"Biochim. Biophys. Acta-Biomembr."},{"key":"ref_65","unstructured":"Mahammad, S., and Parmryd, I. (2015). Methods in Membrane Lipids, Humana Press."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"1029","DOI":"10.1248\/bpb.b16-00047","article-title":"Methyl-beta-Cyclodextrin Impairs the Monocyte-Adhering Ability of Endothelial Cells by Down-Regulating Adhesion Molecules and Caveolae and Reorganizing the Actin Cytoskeleton","volume":"39","author":"Ao","year":"2016","journal-title":"Biol. Pharm. Bull."},{"key":"ref_67","doi-asserted-by":"crossref","unstructured":"Kenworthy, A.K., Schmieder, S.S., Raghunathan, K., Tiwari, A., Wang, T., Kelly, C.V., and Lencer, W.I. (2021). Cholera toxin as a probe for membrane biology. Toxins, 13.","DOI":"10.3390\/toxins13080543"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/0092-8674(95)90404-2","article-title":"Dimerization of cell surface receptors in signal transduction","volume":"80","author":"Heldin","year":"1995","journal-title":"Cell"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"9609","DOI":"10.1073\/pnas.0702668104","article-title":"Chemomechanical mapping of ligand-receptor binding kinetics on cells","volume":"104","author":"Lee","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"1961","DOI":"10.1096\/fasebj.13.14.1961","article-title":"Localization of the insulin receptor in caveolae of adipocyte plasma membrane","volume":"13","author":"Gustavsson","year":"1999","journal-title":"FASEB J."},{"key":"ref_71","doi-asserted-by":"crossref","unstructured":"Care, B.R., and Soula, H.A. (2011). Impact of receptor clustering on ligand binding. BMC Syst. Biol., 5.","DOI":"10.1186\/1752-0509-5-48"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1146\/annurev.cellbio.14.1.111","article-title":"Functions of lipid rafts in biological membranes","volume":"14","author":"Brown","year":"1998","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"18179","DOI":"10.1074\/jbc.RA120.015680","article-title":"BMP-9 and LDL crosstalk regulates ALK-1 endocytosis and LDL transcytosis in endothelial cells","volume":"295","author":"Tao","year":"2020","journal-title":"J. Biol. Chem."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"45035","DOI":"10.1038\/srep45035","article-title":"Dynasore impairs VEGFR2 signalling in an endocytosis-independent manner","volume":"7","author":"Basagiannis","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_75","doi-asserted-by":"crossref","unstructured":"Mironov, A.A., Mironov, A., Sanavio, B., Krol, S., and Beznoussenko, G.V. (2023). Intracellular Membrane Transport in Vascular Endothelial Cells. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24065791"},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1007\/s00418-001-0367-x","article-title":"Vesicle formation and trafficking in endothelial cells and regulation of endothelial barrier function","volume":"117","author":"Minshall","year":"2002","journal-title":"Histochem. Cell Biol."},{"key":"ref_77","doi-asserted-by":"crossref","unstructured":"Larsson, E., Mor\u00e9n, B., McMahon, K.-A., Parton, R.G., and Lundmark, R. (2023). Dynamin2 functions as an accessory protein to reduce the rate of caveola internalization. J. Cell Biol., 222.","DOI":"10.1083\/jcb.202205122"},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1091\/mbc.E20-03-0199","article-title":"ALK1 regulates the internalization of endoglin and the type III TGF-beta receptor","volume":"32","author":"Tazat","year":"2021","journal-title":"Mol. Biol. Cell"},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"791","DOI":"10.1093\/cvr\/cvm097","article-title":"Caveolin-1 interacts and cooperates with the transforming growth factor-beta type I receptor ALK1 in endothelial caveolae","volume":"77","author":"Santibanez","year":"2008","journal-title":"Cardiovasc. Res."},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"4091","DOI":"10.1242\/jcs.188219","article-title":"VEGF induces signalling and angiogenesis by directing VEGFR2 internalisation through macropinocytosis","volume":"129","author":"Basagiannis","year":"2016","journal-title":"J. Cell Sci."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"738","DOI":"10.1038\/cr.2015.60","article-title":"Internalization of the TGF-\u03b2 type I receptor into caveolin-1 and EEA1 double-positive early endosomes","volume":"25","author":"He","year":"2015","journal-title":"Cell Res."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1186\/s12964-015-0102-1","article-title":"Dynasore-not just a dynamin inhibitor","volume":"13","author":"Preta","year":"2015","journal-title":"Cell Commun. Signal."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"1308","DOI":"10.1101\/gad.301549.117","article-title":"DACH1 stimulates shear stress-guided endothelial cell migration and coronary artery growth through the CXCL12\u2013CXCR4 signaling axis","volume":"31","author":"Chang","year":"2017","journal-title":"Genes Dev."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"eabo7958","DOI":"10.1126\/sciadv.abo7958","article-title":"JAG1-NOTCH4 mechanosensing drives atherosclerosis","volume":"8","author":"Souilhol","year":"2022","journal-title":"Sci. Adv."},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1038\/nrm2592","article-title":"Mechanotransduction in development: A growing role for contractility","volume":"10","author":"Wozniak","year":"2009","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"eaar4953","DOI":"10.1126\/scitranslmed.aar4953","article-title":"A BMP\/activin A chimera is superior to native BMPs and induces bone repair in nonhuman primates when delivered in a composite matrix","volume":"11","author":"Seeherman","year":"2019","journal-title":"Sci. Transl. Med."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"1176","DOI":"10.1089\/scd.2011.0293","article-title":"Bone morphogenetic protein-2-induced signaling and osteogenesis is regulated by cell shape, RhoA\/ROCK, and cytoskeletal tension","volume":"21","author":"Wang","year":"2012","journal-title":"Stem Cells Dev."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/13\/3\/285\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T13:55:07Z","timestamp":1760104507000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/13\/3\/285"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,2,5]]},"references-count":87,"journal-issue":{"issue":"3","published-online":{"date-parts":[[2024,2]]}},"alternative-id":["cells13030285"],"URL":"https:\/\/doi.org\/10.3390\/cells13030285","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,2,5]]}}}