E4orf4 triggers repetitive NE rupture/repair and loss of NE integrity in U2OS cells; related to Fig. 7 C. Video microscopy of representative U2OS cells expressing GFP-NLS (gray) and Flag-E4orf4-mCherry (magenta); a cell undergoes repetitive cycles of NE rupture/repair associated with pronounced nuclear deformation (top cell), and another cell shows repetitive NE rupture/repair followed by a prolonged loss of NE integrity that is associated with intense cytoplasmic E4orf4 fluorescence and cell blebbing (bottom cell). Cell imaging was performed for a 10-h period at 5-min intervals using a PerkinElmer UltraVIEW spinning disk confocal equipped with CO2/thermoregulated chamber and 40× 0.75 NA objective; single-plane images are displayed at five frames/s.
