TABLE 2.
Time course of intracellular localization of WT, A53T, and A30P α-Syn
| α-Syn isoform |
Time (hr) | Plasma membrane (%) |
Cytoplasm (%) | Nucleus (%) |
|---|---|---|---|---|
| WT | 0 | 0 | 0 | 0 |
| 2 | 209 (100) | 0 | 0 | |
| 4 | 90 (38) | 148 (62) | 0 | |
| 8 | 4 (18) | 230 (82) | 0 | |
| 12 | 11 (4) | 263 (96) | 0 | |
| A30P | 0 | 0 | 0 | 0 |
| 2 | 0 | 206 (93) | 15 (7) | |
| 4 | 0 | 269 (81) | 64 (19) | |
| 8 | 0 | 251 (82) | 55 (18) | |
| 12 | 0 | 164 (77) | 49 (23) | |
| A53T | 0 | 0 | 0 | 0 |
| 2 | 20 (8) | 238 (92) | 0 | |
| 4 | 38 (15) | 217 (85) | 0 | |
| 8 | 228 (59) | 157 (41) | 0 | |
| 12 | 131 (34) | 250 (66) | 0 |
Y382 cultures were grown to midlog phase in −Trp medium containing 2% sucrose, harvested, and resuspended in −Trp medium containing 2% galactose to induce expression of the indicated GFP fusion proteins. At various times thereafter, an aliquot of cells was removed and scored for subcellular localization of GFP fluorescence. Cells that exhibited clear enhancement of peripheral fluorescence were scored as localizing to the plasma membrane; otherwise, they were scored as localizing to the cytoplasm if diffusely fluorescent or to the nucleus if they exhibited enhanced nuclear fluorescence. For each time point, multiple random fields were counted.