Table I.
Calculated parameters derived from FCS measurements
Proteins | Nucleoplasm | Splicing factor compartments | ||||||
τDa | Dfa | τDb | Dfb | τDa | Dfa | τDb | Dfb | |
ms | µm2s−1 | ms | µm2s−1 | ms | µm2s−1 | ms | µm2s−1 | |
U1-70Ka | 0.56 ± 0.08 | 28.21 ± 4.22 | 24.92 ± 3.08 | 0.63 ± 0.08 | 0.67 ± 0.17 | 24.98 ± 7.41 | 33.68 ± 3.77 | 0.47 ± 0.05 |
U2A′a | 0.67 ± 0.18 | 24.85 ± 6.87 | 19.91 ± 4.68 | 0.82 ± 0.20 | 0.72 ± 0.11 | 22.30 ± 3.70 | 26.82 ± 5.98 | 0.61 ± 0.15 |
hPrp31a | 0.99 ± 0.23 | 16.74 ± 5.40 | 18.08 ± 3.13 | 0.88 ± 0.15 | 0.97 ± 0.12 | 16.27 ± 2.01 | 20.55 ± 4.49 | 0.79 ± 0.18 |
hPrp4a | 0.76 ± 0.17 | 21.35 ± 4.49 | 24.68 ± 3.03 | 0.64 ± 0.08 | 1.06 ± 0.35 | 15.96 ± 4.54 | 22.46 ± 2.83 | 0.71 ± 0.10 |
hPrp8a | 0.64 ± 0.12 | 24.97 ± 4.50 | 30.29 ± 1.44 | 0.52 ± 0.02 | 0.78 ± 0.16 | 20.79 ± 4.34 | 33.83 ± 4.91 | 0.47 ± 0.07 |
Snu114a | 0.76 ± 0.24 | 22.09 ± 5.83 | 26.45 ± 5.16 | 0.61 ± 0.12 | 1.01 ± 0.09 | 15.58 ± 1.47 | 27.18 ± 2.78 | 0.58 ± 0.06 |
U1-70KΔ1–197 | 0.58 ± 0.19 | 28.81 ± 6.36 | 6.80 ± 1.50 | 2.40 ± 0.55 | 0.82 ± 0.10 | 19.39 ± 6.36 | 6.18 ± 1.89 | 2.70 ± 0.66 |
EGFP | 0.56 ± 0.05 | 27.87 ± 2.22 | NA | NA | ND | ND | ND | ND |
NA, not applicable. Dynamics of snRNP proteins were measured in the nucleoplasm and the splicing factor compartments by FCS. The delay time values τDa for EGFP are comparable with the values of the fast component detected in the FCS measurements of snRNP proteins. The values for the delay times τDb describe the diffusion of the slow-moving component that reflects snRNP complexes. τDa and τDb were used to calculate the diffusion coefficients Dfa and Dfb, respectively. The mean ± SD from 8–10 cells is shown.
BAC stable cell line.