Using fluorescence resonance energy transfer-based biosensors to probe Rho GTPase activation during phagocytosis

8Citations
Citations of this article
23Readers
Mendeley users who have this article in their library.
Get full text

Abstract

The p21-family members of Rho GTPases are important for the control of actin cytoskeleton dynamics, and are critical regulators of phagocytosis. The three-dimensional structure of phagosomes and the highly compartmentalized nature of the signaling mechanisms during phagocytosis require high-resolution imaging using ratiometric biosensors to decipher Rho GTPase activities regulating phagosome formation and function. Here we describe methods for the expression and ratiometric imaging of FRET-based Rho GTPase biosensors in macrophages during phagocytosis. As an example, we show Cdc42 activity at the phagosome over Z-serial planes. In addition, we demonstrate the usage of a new, fast, and user-friendly deconvolution package that delivers significant improvements in the attainable details of Rho GTPase activity in phagosome structures.

Cite

CITATION STYLE

APA

Miskolci, V., Hodgson, L., & Cox, D. (2017). Using fluorescence resonance energy transfer-based biosensors to probe Rho GTPase activation during phagocytosis. In Methods in Molecular Biology (Vol. 1519, pp. 125–143). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6581-6_9

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free