Table 4.
Effect of siRNA treatment on PrPC endocytosis rate and cell-surface expression
PrPC cell-surface expression, influence of siRNA treatment on PrPC cell-surface expression. Cells were incubated for 3 days with siRNAs. PrPC cell-surface level was detected with the POM2-Eu/POM2-APC antibody pair. PrPC cell-surface expression of siRNA-treated cells is expressed as -fold untreated cells. PrPC endocytosis, influence of siRNA treatment on the endocytosis rate of PrPC. Cells were incubated for 3 days with siRNAs and labeled with POM2-Eu at 4 °C. To induce endocytosis, cells were exposed to 37 °C. At time point t0 = 0 min and t1 = 30 min POM2-APC was added. The endocytosis rate was calculated as described under “Experimental procedures.”
| siRNA | PrPC cell-surface expression |
PrPC endocytosis |
||
|---|---|---|---|---|
| Mean ± S.D. (t0) | Normalized to untreated (t0) | Mean ± S.D. (t0—t30) | Endocytosis rate | |
| Untreated | 448.43 ± 103.37 | 1 ± 0.23 | 213.62 ± 110.26 | 0.48 ± 0.36 |
| PrP | 229.24 ± 108.70 | 0.51 ± 0.24 | 52.65 ± 118.97 | 0.23 ± 0.63 |
| AP2M1 | 443.52 ± 19.41 | 0.99 ± 0.04 | 40.64 ± 30.35 | 0.09 ± 0.07 |
| EHD1 | 493.18 ± 28.72 | 1.10 ± 0.06 | 254.44 ± 80.13 | 0.52 ± 0.19 |
| GIT2 | 459.97 ± 50.50 | 1.03 ± 0.11 | 228.70 ± 90.94 | 0.50 ± 0.25 |
| M6PR | 435.74 ± 82.25 | 0.97 ± 0.18 | 79.65 ± 102.08 | 0.18 ± 0.27 |
| RAB5A | 202.00 ± 89.66 | 0.45 ± 0.20 | −51.60 ± 115.41 | −0.26 ± 0.46 |
| VPS28 | 349.60 ± 125.23 | 0.78 ± 0.28 | 199.25 ± 136.61 | 0.57 ± 0.59 |
| VPS35 | 344.96 ± 122.54 | 0.77 ± 0.27 | 65.19 ± 137.30 | 0.19 ± 0.47 |