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Radeva, Mariya Y.; Waschke, Jens ORCID: 0000-0003-1182-5422; Fuchs, Michael (2026): STED/AFM as a tool to investigate mechanical and adhesive properties of migrating keratinocytes. Cell Adhesion & Migration, 20 (1): 2657675. ISSN 1933-6918

[thumbnail of STED_AFM_as_a_tool_to_investigate_mechanical_and_adhesive_properties_of_migrating_keratinocytes.pdf] Creative Commons Namensnennung (CC BY)
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STED_AFM_as_a_tool_to_investigate_mechanical_and_adhesive_properties_of_migrating_keratinocytes.pdf

Abstract

E-cadherin is a key component of adherens junctions which maintains epithelial integrity. In keratinocytes, wound healing requires dynamic modulation of adhesion and cytoskeletal organization. Using a wound healing assay combined with stimulated emission depletion/atomic force microscopy (STED/AFM), we analysed E-cadherin binding during murine keratinocyte migration. Wound closure occurred within 6 h and was accompanied by E-cadherin accumulation at the leading edge. Transient expression of E-Cadherin-SNAP enabled investigation of E-cadherin interactions. Inhibition of actin polymerization abolished E-cadherin binding and reduced cellular stiffness. Imaging of SiR-actin-labeled cells enabled simultaneous visualization of migration and measurement of binding and mechanical properties. E-cadherin retained functional binding properties during migration. These findings establish STED/AFM as a powerful method to investigate single-molecule binding properties in migrating cells.

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