Quantification of polarity of dendritic and axonal proteins in control and PKD1-KD-transfected neurons
| Protein | Number of cells analyzed (n) | Polarity index | Fluorescence intensity |
|---|---|---|---|
| TfR-GFP | |||
| Control (nontransfected) | 25 | 10.75 ± 2.5 | 2930 ± 28 |
| PKD1-WT | 25 | 9.25 ± 0.5 | 2927 ± 21 |
| PKD1-WT (S916A) | 25 | 4.11 ± 0.2* | 2950 ± 45 |
| PKD1-KD | 25 | 1.11 ± 0.1* | 2842 ± 86 |
| PKD1-KD (P155 G) | 25 | 10.60 ± 1.8 | 3045 ± 65 |
| PKD1-KD (S916E) | 25 | 5.47 ± 0.1* | 3028 ± 62 |
| LRP (endogenous) | |||
| Control (nontransfected) | 25 | 10.6 ± 2.2 | 864 ± 48 |
| PKD1-KD | 25 | 1.38 ± 0.3* | 935 ± 20 |
| PKD1-KD (P155 G) | 25 | 8.8 ± 1.4 | 880 ± 40 |
| shRNA PKD1 | 25 | 1.45 ± 0.5* | 1025 ± 65 |
| Scrambled-shRNA PKD1 | 25 | 10.8 ± 0.6 | 940 ± 30 |
| GFP-VAMP2 | |||
| Control (nontransfected) | 20 | 1.25 ± 0.2 | 2450 ± 110 |
| PKD1-KD | 20 | 1.36 ± 0.4 | 2480 ± 80 |
| L1-YFP | |||
| Control (nontransfected) | 20 | 1.15 ± 0.2 | 860 ± 20 |
| PKD1-KD | 20 | 1.20 ± 0.7 | 886 ± 24 |
| MAP2 | |||
| Control (nontransfected) | 42 | 16.9 ± 0.8 | 1250 ± 15 |
| PKD1-KD | 42 | 18.2 ± 0.6 | 1265 ± 35 |
| Tau | |||
| Control (nontransfected) | 36 | 0.17 ± 0.04 | 466 ± 24 |
| PKD1-KD | 36 | 0.13 ± 0.03 | 480 ± 42 |
Values are mean ± SEM.
↵*Significantly different from the corresponding control group (p < 0.05). Polarity index was calculated as described in Materials and Methods by dividing the dendritic by the axonal average fluorescence intensity expressed in pixels. A nonpolarized distribution gives a value close to 1. Fluorescence intensity values are expressed in pixels; 14 bit images were used for this type of measurement. Fluorescent intensity measurements of TfR-GFP, endogenous LRP, GFP-VAMP2, L1-YFP, MAP2, or Tau were performed in the cell body of control and transfected neurons. At least 20 cells from three different cultures were analyzed for each experimental condition.