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Files in this Data Supplement:
Movie 1. Mature PA-GFP-Sm-tagged snRNPs show unrestricted exchange between nuclear structures. Repeated activation of a region of interest containing a CB results in rapid accumulation of PA-GFP-SmB in speckles throughout the entire nucleus. Bar, 22 μm.
Movie 2. Mature PA-GFP-Sm-tagged snRNPs show unrestricted exchange between nuclear structures. Repeated activation of a region of interest containing a CB results in rapid accumulation of PA-GFP-SmB in both speckles and CBs throughout the entire nucleus. Bar, 22 μm.
Movie 3. Mature PA-GFP-Sm-tagged snRNPs show unrestricted exchange between nuclear structures. Repeated activation of a region of interest containing a speckle results in rapid accumulation of PA-GFP-SmB in both speckles and CBs throughout the entire nucleus. Bar, 22 μm.
Movie 4. Mature snRNPs do not show a measurable export from the nucleus to the cytoplasm. HeLa cells stably expressing YFP-SmB were bleached repeatedly using a region of interest including a region of the cytoplasm of one cell and a region of the nucleus of its neighbour. The cells were imaged following each bleach event. In the cell undergoing bleaching of the nucleus, the entire nucleus was rapidly depleted of YFP-SmB. By contrast, the cell undergoing cytoplasmic bleaching lost virtually none of its nuclear signal. Bar, 22 μm.
Movie 5. Recently imported snRNPs are unable to accumulate in speckles. Repeated activation of a region containing a CB results in rapid accumulation of signal in distant CBs. No accumulation in speckles can be detected. Bar, 22 μm.
Movie 6. At early timepoints, YFP-SmB can exit the nucleus into the cytoplasm. HeLa cells expressing YFP-SmB for 24 hours were bleached repeatedly using a region of interest including a region of the cytoplasm of one cell and a region of the nucleus of its neighbour. The cells were imaged following each bleach event. In the cell undergoing bleaching of the nucleus, the entire nucleus was rapidly depleted of YFP-SmB. In the cell undergoing cytoplasmic bleaching, the nucleus also lost a significant amount of signal. Bar, 22 μm.
Movie 7. At early timepoints, YFP-SmB can exit the nucleus into the cytoplasm. HeLa cells expressing YFP-SmB for 24 hours were bleached repeatedly using a region of interest including a region of the cytoplasm of one cell and a region of the nucleus of its neighbour. The cells were imaged following each bleach event. In the cell undergoing bleaching of the nucleus, the entire nucleus was rapidly depleted of YFP-SmB. In the cell undergoing cytoplasmic bleaching, the nucleus also lost a significant amount of signal. Bar, 22 μm.
Movie 8. LMB does not affect the ability of speckles to accumulate snRNPs. HeLa cells expressing PA-GFP-SmB were treated with LMB for 4 hours. A region of the nucleus containing a CB was repeatedly activated with a 406 nm laser and short time-lapse sequences were taken between each bleach event to monitor the spread of PA-GFP-SmB in the nucleus. The signal rapidly accumulated in speckles throughout the nucleus. Bar, 11 μm.
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