Fig. 1

Nuclear protrusion and herniation, mitotic failure, and aberrant cytokinesis in lamin A/C-suppressed HOSE cells. Lamin A and C were suppressed by siRNA (Santa Cruz biotechnology Inc, Santa Cruz, CA) in primary human ovarian epithelial (HOSE) cells, using lipofectamine 2000 according to the manufacturer protocol (Invitrogen, CA). a At day 3, the lamin A/C-suppressed cells were analyzed using immunofluorescence microscopy. An example of a cell undergoing nuclear protrusion and herniation is shown (arrow), for nuclear staining with DAPI (blue) and lamin A/C (red). b The lamin A/C-suppressed primary HOSE cells that were previously transfected with GFP-histone H2B were monitored for nuclear changes by time-lapse video fluorescence microscopy over a 48-h period. Time-lapse video microscopy was performed 12 h after lamin A/C suppression. Cells were seeded in 24 well falcon plate and transfected the next day with scramble siRNA (control) or lamin A/C siRNA in serum reduced Opti-MEM media. Image acquisition was performed every 5 min for 48 h with a 40× dry objective lens on Nikon Eclipse TE 300 microscope linked to a Roper Scientific photometrics 12-bit range Camera using Meta imaging series (MetaVue) software. Stacked images were assembled with MetaVue software to make the movies. Two examples of sequential time-lapse images 15 min apart are shown. Arrows indicate the nuclear herniation and the formation of micronuclei, which gradually faded. Presumably, aneuploidy was resulted in the remaining nuclei. c Sequential time-lapse images (images frame #1 to 12) 15 min apart are shown for a 3-h segment of a video of a mitotic failure of a dividing cell. Arrows indicate the nuclei first underwent DNA condensation, separation, and then fused back to form presumably tetraploid nuclei. d Sequential time-lapse images (frame #1 to 8) 15 min apart show an aberrant mitotic process. Arrows indicate the nuclei undergoing a tripolar division. Likely, aneuploid cells were formed