Movie 5.

PIP-FUCCI accurately reports cell cycle phases in H1299 cells. Video of a dividing H1299-FUCCI cell over 24 h (20 min intervals), compiled from a single Z-plane captured every 20 min. PIP-FUCCI uses regulatory domains PCNA interacting protein degron from human Cdt11-17 (PIP) (mVenus, green) and Geminin1-110 (mCherry, magenta) to differentially express fluorescent proteins during cell cycle phases. Labels appear as the cell transitions to each phase (G1, S, G2, M, G1), scale bar: 20 μm. PIP-mVenus accumulates in the nucleus during G1 and is rapidly lost during onset of S phase, where mCherry-Geminin begins to rise and is expressed throughout S, G2, and M phases. During the S-G2 transition, mVenus accumulates again, and both mVenus and mCherry are co-expressed until division. M phase is marked by nuclear envelope breakdown and diffusion of mVenus and mCherry signals throughout the cell. Following cytokinesis, only mVenus is expressed in the two daughter cell nuclei, marking G1. Video is the same cell as Fig. 6C.