Visualization of Radiation-Induced Cell Cycle Kinetics with a Fluorescent Ubiquitination-Based Cell Cycle Indicator (Fucci).

ABSTRACT

Among the methods for detecting cell cycle kinetics in tumor cells, fluorescent ubiquitination-based cell cycle indicator (Fucci) is innovative because it allows observation in live cells without losing spatiotemporal information. We succeeded in using the Fucci system to visualize radiation-induced G2 arrest in tumor cells with deficient p53 function. Here we describe protocols for establishing Fucci-expressing cell lines and analyzing radiation-induced G2 arrest kinetics in three different models monolayer cell cultures, spheroids, and xenografted solid tumors in mice.