Visualizing Vpr-induced cell cycle arrest and cell death.

<p>HeLa/Fucci2 cells were transfected with pME18Neo/Flag-Vpr-IRES-ECFP or the control pME18Neo/Flag-IRES-ECFP. At 24 h after transfection, cells were monitored by time-lapse imaging using LCV110 Imaging System at 15 minute intervals until 96 h post-transfection. Cells showing red or yellow fluorescence in the nucleus are in the G1 or S/G2/M phases of the cell cycle, respectively. ECFP and Fucci2 fluorescence is shown at 24, 36, 48, 60, 72, 84, and 96 h post-transfection. Eight ECFP-positive cells were selected as representative images, and are marked as *1 to *8 on the right of each cell. The scale bars represent 100 μ. High-magnification images of ECFP-expressing cells *1 to *8 are shown for cells transfected with the control pME18Neo/Flag-IRES-ECFP (*1 and *2) at 24, 27, 28, 36, 42, 43, 48, 60, 67, 72, 80, 88, or 96 h, or pME18Neo/Flag-Vpr-IRES-ECFP (*3 to *8) at 24, 26, 28, 36, 48, 54, 60, 66, 72, 73, 84, 91, or 96 h after transfection.