Fig. 7. Further localisation of emerin and lamin A/C by representing z-sections as x-z slices through cells. A series of z-sections (0.2 µm apart) were taken through the nuclei of COS-7 cells transfected with either wild-type or Del236-241 EGFP-emerin. The z-sections shown are for: (A) a cell transfected with wild-type EGFP-emerin (18 z-sections taken in total) 28 hours after being released from nocodazole; (B) a cell transfected with Del236-241 EGFP-emerin (31 z-sections taken in total) 28 hours after being released from nocodazole; and (C) a cell transfected with Del236-241 EGFP-emerin (25 z-sections taken in total) 35 hours after being released from nocodazole treatment. All z-sections were collected and represented as x-z slices through the cells. The projections of all the z-sections taken show emerin in green, lamin A/C in red and colocalisation in yellow. The white lines show where the z-sections were collected: the centre of the nucleus 0, 0+5 µm and 0-5 µm. The line thickness was ±0.3 µm for wild-type cells at 28 hours, ±0.3 µm for Del236-241 at 28 hours and ±0.6 µm for Del236-241 at 35 hours. Each x-z slice is shown separately in black and white for clarity. Cells expressing Del236-241 EGFP-emerin 28 and 35 hours after synchronisation were microscopically similar. The correct localisation of emerin and lamin A/C to the nuclear membrane is shown to be important, as the mutant form of emerin (Del236-241) redistributes lamin A/C with resultant defects in the nuclear architecture. Radiance Confocal Laser Scanning System; bar, 5 µm.
