Protocol to quantify desmoglein 3 tension at cell-cell adhesions using FRET-based tension sensors
Year: 2026
Type: article
Abstract: 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
Source: STAR Protocols
Authors: Bahareh Tajvidi Safa, Timothy Goldsmith, Zachary Kezhaya, Sarah Seman, Daniel E. Conway, Ruiguo Yang
Language: English
Subfield: Cell Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,550
Awards: G1130520335, G1149519361, CAREER: Characterization of the Strain Rate-Dependent Mechanical Behavior of the Cell-Cell Adhesion Interface, G1707597437, CAREER: Characterization of the Strain Rate-Dependent Mechanical Behavior of the Cell-Cell Adhesion Interface, G2228770426, Measurement of Mechanical Tension Across Desmosomes, Probing the Active Tensioning Response Under Sustained Strain in Epithelial Cells, G66062016, G9000066001, Direct and Quantitative Probing of Desmosome Mechanotransduction
OpenAlex ID: W7164925741