Epithelial cell spreading assay on E-cadherin-coated glass or PDMS substrates for microscopy-based analysis of cadherin adhesions.

Lilian Schimmel, Ivar Noordstra

STAR protocols 2023 Dec 15

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Adherens junctions (AJs) are multi-protein adhesion structures that couple contractile actomyosin networks of epithelial cells within a tissue. Here, we present an epithelial cell spreading assay on E-cadherin-coated glass or polydimethylsiloxane (PDMS) substrates for detailed microscopy-based analysis of cadherin adhesions. We describe steps for preparation of glass coverslips and PDMS gels, E-cadherin coating, and epithelial cell spreading. Epithelial cells can be seeded on E-cadherin-coated surfaces, thereby mimicking AJ formation in X-Y dimension, making it suitable for microscopy analysis. For complete details on the use and execution of this protocol, please refer to Noordstra et al. (2023).1. Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.

Citation

Lilian Schimmel, Ivar Noordstra. Epithelial cell spreading assay on E-cadherin-coated glass or PDMS substrates for microscopy-based analysis of cadherin adhesions. STAR protocols. 2023 Dec 15;4(4):102626