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Figure 6


Fig. 6. Stability of the nuclear lamina following modification by RNAi. HeLa cells were co-transfected with RNAi vectors and vector expressing the relevant GFP-laminA/C or B1 to monitor dynamics. After 48 hours, cells with low intensity reporter expression were selected and fluorescence recovery after photobleaching monitored. Panels (A) show a typical FRAP profile (from left to right): pre-bleach image; image immediately after bleaching – in each cell analysed two 2 µm zones were bleached, one is shown boxed; and two frames (15 and 60 minutes) from a time-lapse series with images taken at 15 minute intervals over 2 hours. Analysis of bleach zones (B) was used to estimate t1/2 for fluorescent recovery for cells transfected with empty RNAi vector (Control) and cells with lamin B1 (LB1 KD) and lamin A/C (LA/C KD) depleted. Data points shown in B are average intensities of ten bleach zones (five cells) for the long (300 minute) timecourse and 20 bleach zones (ten cells) for the short (50 minute) timecourse. Scale bars: 5 µm.





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