Table I. The lateral mobility of GFP-tagged fusion proteins stably expressed in M3/18 or transiently expressed in tsBN7 cells.
| Cell line | Construct | Growth temperature | Treatment | Diffusion constant (D eff), μm2/s ± SD |
Mobile fraction (Mr), % ± SD |
n |
|---|---|---|---|---|---|---|
| M3/18 | GFP–Dad1 | 39.5°C | – | 0.049 ± 0.010 | 87.9 ± 7.4 | 18 |
| Cycloheximide | 0.044 ± 0.013 | 89.6 ± 8.1 | 8 | |||
| NaCl | 0.251 ± 0.071 | 94.5 ± 5.6 | 5 | |||
| Puromycin | 0.131 ± 0.058 | 89.4 ± 5.7 | 25 | |||
| Tunicamycin | 0.048 ± 0.016 | 89.1 ± 2.5 | 10 | |||
| tsBN7 | LBR–GFP | 34°C | – | 0.331 ± 0.054 | 99.6 ± 5.2 | 4 |
| NaCl | 0.328 ± 0.032 | 97.8 ± 1.3 | 9 | |||
| Puromycin | 0.335 ± 0.029 | 96.3 ± 3.6 | 7 | |||
| 39.5°C | – | 0.331 ± 0.031 | 89.6 ± 2.9 | 8 | ||
| Dad1–GFP | 34°C | – | 0.315 ± 0.064 | 93.3 ± 5.2 | 7 | |
| 39.5°C | – | NA | NA | 15 | ||
| 34°C | – | 0.178 ± 0.037 | 89.5 ± 5.9 | 10 | ||
| GFP–Dad1 | 39.5°C | – | 0.140 ± 0.024 | 88.9 ± 1.9 | 10 |
Means ± SD denotes standard deviation of values for D eff and M f for recovery of fluorescence after photobleaching. The cells were grown overnight in glass-bottomed dishes for cell culture and pretreated as indicated with cycloheximide (80 μM for 30 min), NaCl (an extra 150 mM for 10 min), puromycin (100 μM for 15 min), or tunicamycin (1 μg/ml for 4 h) before the FRAP experiments. NA denotes that no reliable data were obtained (see Results for details).