Figure 1. PIP-FUCCI Lentivirus reports cell cycle status in HUVEC.
(A) PIP-FUCCI lentivirus construct. NLS, nuclear localization signal; HA, HA tag; P2A, self-cleaving peptide 2A; Gem, geminin. (B) Representative time-lapse images of PIP-FUCCI transduced HUVEC, showing mVenus-Cdt11–17 (green) and mCherry-Geminin1–110 (red) expression hourly from end of cytokinesis (M) through next cytokinesis. Scale bar, 20 μm. (C) Quantification of PIP-FUCCI fluorescence intensity/time from cell in B. (D) Average time spent in each cell cycle state (hr). (n=30 cells per phase from 3 replicate movies) (E). Total endothelial cell cycle length, time between mitoses measured by H2B-CFP. (n=15 cells from 3 replicate movies).