Table 1.
FRAP 1 Summary of T1/2 and retention of APC-FL and APC1-1309 at the centrosome.
| APC +/− Treatments | Mobile Pool 2 (%) | Retained Pool 2 (%) | T1/2 Fast Pool 3 (s) | T1/2 Slow Pool (s) |
|---|---|---|---|---|
| APC-FL | 86.23 | 13.77 | 2.77 | 25.58 |
| APC-FL+ noco | 97.30 | 2.70 | 6.79 (p < 0.0001) | 42.67 |
| APC1-1309 | 91.31 | 8.69 | 3.09 | 27.82 |
| APC1-1309+noco | 90.39 | 9.61 | 5.99 (p = 0.045) | 25.79 |
| APC1309 +control siRNA |
87.81 | 12.19 | 3.02 | 23.34 |
| APC1309 + γ-tub siRNA |
83.77 | 16.23 | 5.44 (p = 0.019) | 10.52 |
1 The analysis of fluorescence recovery of GFP-tagged APC-FL and APC1-1309 at the centrosome in HeLa cells (see Figure 2 and Figure 3) was measured using a combination of Excel and algorithms from the GraphPad Prism 5 program. Given the period of recovery measured (100 s), we could only reliably measure levels of significance for differences in the fast phase pool (1–40 s), our major focus in this study. The Table shows values for centrosome retention and recovery rates of fast and slow populations with and without treatment. All fluorescence recovery curves were fitted to a two-phase recovery curve producing two T1/2 values (fast and slow). See Methods for further details. 2 Mobile/retained pools: plateaus reached on the curves were used to determine (or predict in mostcases since recovery curves had not always reached plateau within the time period of analysis) the distribution of mobile and retained pools measured by the % of maximum fluorescence recovery by the Graph Pad Prism 5 program. 3 Rate of recovery: The Table indicates the speed of recovery of fluorescence for the two populations of APC at the centrosome. The fast pool T1/2 values were based on the recovery speed during the first 40 s (T = 0–40 s). The slow pool data shown is an extrapolation and included only for reference.