Effects of DLK–JNK pathway inactivation and Taxol treatment on the different stages of neuronal polarization
| Treatments | na | Stage 1/stage 2b (SD) | p valuec | Stage 2/stage 3d (SD) | p valuec |
|---|---|---|---|---|---|
| NC | 7 | 100.0 (0.5) | 100.0 (5.6) | ||
| DLK RNAi | 7 | 94.7 (2.2) | 0.00004 | 48.8 (8.9) | 0.00000002 |
| DLK RNAi + Taxol | 7 | 86.5 (2.6) | 0.00000001 | 87.4 (2.8) | 0.0002 |
| 0.00003e | 0.0000001e | ||||
| NC + Taxol | 7 | 96.6 (2.0) | 0.0008e | 109.2 (3.5) | 0.003e |
| DMSO | 3 | 100.0 (0.6) | 100.0 (5.2) | ||
| SP600125 | 3 | 98.0 (1.2) | 0.055 (NS) | 52.0 (4.2) | 0.0002 |
| SP600125 + Taxol | 3 | 85.0 (4.2) | 0.003 | 89.6 (6.1) | 0.087 (NS) |
| 0.006e | 0.0009e | ||||
| DMSO + Taxol | 3 | 95.0 (2.1) | 0.016e | 108.1 (5.1) | 0.13 (NS)e |
| NC | 4 | 100.0 (1.4) | 100.0 (2.0) | ||
| DLK RNAi | 4 | 93.9 (4.3) | 0.036 | 43.0 (11.1) | 0.00006 |
| DLK RNAi + DLK | 4 | 100.0 (0.82) | 1.0 (NS) | 92.2 (7.0) | 0.076 (NS) |
| 0.032f | 0.0003f | ||||
| DLK RNAi + DLK/KN | 4 | 91.9 (6.9) | 0.060 (NS) | 26.4 (11.7) | 0.00002 |
| 0.63 (NS)f | 0.086 (NS)f | ||||
| NC | 4 | 100.0 (0.5) | 100.0 (3.4) | ||
| DLK RNAi | 4 | 94.5 (3.9) | 0.030 | 57.9 (5.1) | 0.000009 |
| DLK RNAi + JNK1-MKK7 | 4 | 99.0 (2.1) | 0.38 (NS) | 96.0 (4.6) | 0.21 (NS) |
| 0.086 (NS)f | 0.00003f | ||||
| DLK RNAi + JNK1/KN-MKK7/KN | 4 | 93.2 (6.4) | 0.078 (NS) | 57.8 (7.2) | 0.00002 |
| 0.75 (NS)f | 0.29 (NS)f |
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↵aNumber of independent experiments performed on different days using freshly prepared primary neurons.
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↵bThe efficiency of stage 1/stage 2 transitions was evaluated by calculating the relative number of stage 2 and stage 3 neurons against the total number of neurons at 3 DIV. Indicated values represent the efficiency of stage transitions in treated cells relative to that of control cells (percentage).
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↵cThe efficiency of stage 1/stage 2 transitions or stage 2/stage 3 transitions was compared between control cells (NC, nonsilencing control; DMSO, vehicle) and shRNA vector (DLK#1)-transfected or JNK inhibitor (10 μm SP600125)-treated cells, using two-tailed Student's t test; p < 0.05 was considered significant. NS, Not significant.
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↵dThe efficiency of stage 2/stage 3 transitions was evaluated by calculating the relative number of stage 3 neurons against the number of stage 2 and stage 3 neurons at 3 DIV. Indicated values represent the efficiency of stage transitions in treated cells relative to that of control cells.
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↵ep values were obtained by comparison with cells that were not treated with Taxol. Taxol (5 nm) was added at 1 DIV.
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↵fp values were obtained by comparison with DLK RNAi cells.
