Figure 5
FRAP analysis of GFP-NSD3S shows that NSD3S is a dynamic component of heterochromatin. (A) Cellular localization of NSD3S. GFP-NSD3S expressing L929 cells were fixed with paraformaldehyde and localization of fusion protein was visualized with fluorescence microscopy. Dense foci seen by Hoechst staining represent pericentric heterochromatin regions (a). GFP-NSD3S labelling co-localizes at these sites (b). (B) Time-lapse serie of confocal images of living cells expressing GFP-NSD3S during a FRAP experiment. Images were recorded before (Prebleach) and at different time intervals after the bleach. Arrows indicate the photobleached areas. (C) Parameters of NSD3S protein dynamics derived from FRAP curves. The half-time of fluorescence recovery (t1/2), the percentage of fluorescence recovery after 30 s, and the number of FRAP curves used are indicated. (D) Representation of the number of cells characterized by a given t1/2 value after cell synchronization by serum starvation. (E) Representation of the number of cells characterized by a given t1/2 value after cell synchronization by nocodazole treatment.