Protocol to quantify desmoglein 3 tension at cell-cell adhesions using FRET-based tension sensors
Desmoglein 3 (Dsg3) is a crucial component of cell-cell junctions, contributing to mechanotransduction and maintaining tissue integrity. Here, we present a protocol to quantify tension at keratinocyte cell-cell adhesion sites using a Dsg3 fluorescence resonance energy transfer (FRET)-based tension sensor. We describe steps for increasing viral titer by amplifying adenovirus in HEK293A cells and optimizing infection conditions to ensure uniform expression of FRET signals. We detail procedures for live-cell imaging of keratinocytes and image analysis to quantify the FRET ratio. For complete details on the use and execution of this protocol, please refer to Jin et al. 1
Authors
- Daniel E. Conway (ORCID: https://orcid.org/0000-0001-6624-177X)
- Ruiguo Yang (ORCID: https://orcid.org/0000-0002-1361-4277)
- Bahareh Tajvidi Safa (ORCID: https://orcid.org/0000-0003-1280-5319)
- Timothy Goldsmith
- Zachary Kezhaya
- Sarah Seman
Institutions
- The Ohio State University Comprehensive Cancer Center – Arthur G. James Cancer Hospital and Richard J. Solove Research Institute (US)
- Quantitative BioSciences (US)
- The Ohio State University (US)
- Michigan State University (US)
Publication Details
- Journal
- STAR Protocols
- Published
- 2026-06-16
- DOI
- https://doi.org/10.1016/j.xpro.2026.104630
- Primary Topic
- Cellular Mechanics and Interactions
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- National Science Foundation
- National Institute of General Medical Sciences
- National Institute of Arthritis and Musculoskeletal and Skin Diseases