Protocol for quantitative live-cell imaging of early GPCR trafficking using acid-stable afCFP-Venus FRET
Here, we present a protocol to visualize and quantify β -arrestin-dependent signaling of the angiotensin II type 1 receptor (AT1R). We describe steps for detecting receptor- β -arrestin interactions during early trafficking events under our imaging conditions using an afCFP-Venus FRET pair. We then detail procedures for visualizing early intracellular signaling dynamics following AT1R activation and for analyzing GPCR trafficking and signaling dynamics in living cells.
Authors
- Yoichi Takami (ORCID: https://orcid.org/0000-0001-9018-6707)
- Akitoshi Hara (ORCID: https://orcid.org/0000-0002-9369-3293)
- Toshimasa Takahashi (ORCID: https://orcid.org/0000-0002-5203-155X)
- Koichi Yamamoto (ORCID: https://orcid.org/0009-0000-0918-1088)
- W.T. Liu (ORCID: https://orcid.org/0000-0003-4885-1675)
- Lisha Yi
- Nanxiang Yin
- Heedong Jeon
- Cheng Wang
- Yu Guo
- Daisuke Nawa
- Ziwei Wang
Institutions
- University of Toronto (CA)
- The University of Osaka (JP)
Publication Details
- Journal
- STAR Protocols
- Published
- 2026-06-30
- DOI
- https://doi.org/10.1016/j.xpro.2026.104664
- Primary Topic
- Receptor Mechanisms and Signaling
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Japan Society for the Promotion of Science