Summary of the effects of hKID, KIF4A, and hKID/KIF4A RNAi in human cells
| Parameter\Treatment | Control RNAi | hKID RNAi | KIF4A RNAi | hKID/KIF4A RNAi |
| Mitotic index (MI) (mean ± SEM) | 6.1 ± 0.1% | 7.3 ± 0.3% (*) | 6.7 ± 0.3% | 17.9 ± 1.2% (*) |
| Length of mitosis (median; Q1; Q3) | 35; 30; 40 min | 40; 35; 60 min | 40; 30; 60 min | 90; 65; 150 min (***) |
| Prometaphases (mean ± SEM) | 30.7 ± 3.2% | 42.5 ± 1.6% (*) | 38.4 ± 5.7% | 78.4 ± 3.9% (***) |
| Lagging chromosomes (mean ± SEM) | 3.19 ± 0.52% | 10.77 ± 3.4% | 11.69 ± 2.05% | 25.7 ± 2.25% (*) |
| Cytokinesis failures (mean ± SEM) | 2.37 ± 1.1% | 1.1 ± 0.4% | 4.85 ± 1.8% | 10.45 ± 0.41% |
| ≥3 uncongressed chromosomes | 8.8% | 19.7% | 18.0% | 62.5% |
| Cell death (mean ± SD) | 1.85 ± 0.31% | 2.23 ± 0.26% | 3.06 ± 0.25% (*) | 4.09 ± 0.75% |
| Bipolar spindle size (mean ± SD) | 8.70 ± 0.68 µm | 7.77 ± 0.64 µm (***) | 8.53 ± 0.73 µm | 9.94 ± 1.36 µm (***) |
| Monopolar spindles (mean ± SEM) | 3.98 ± 0.08 µm | 3.12 ± 0.11 µm (***) | 3.78 ± 0.06 µm | 2.85 ± 0.06 µm (***) |
| IK distance (mean ± SD) | 1.01 ± 0.08 µm | 1.00 ± 0.07 µm | 1.27 ± 0.09 µm (***) | 1.24 ± 0.10 µm (***) |
| Oscillation half period (τx) | 52.5 s | 52.5 s | 60 s | 75 s |
| Oscillation speed (mean ± SD) | 1.23 ± 0.21 µm/min | 1.13 ± 0.18 µm/min (**) | 1.13 ± 0.21 µm/min (*) | 1.12 ± 0.17 µm/min (**) |
| Nocodazole sens. MI (mean ± SEM) | 9.9 ± 0.8% (*) | 22.7 ± 1.1% (**) | 4.5 ± 0.3% | 31.9 ± 1.4% (**) |
| MT density (EB1-GFP)(mean ± SEM) | 17.7 ± 0.01 | 14.6 ± 0.03 | 23.5 ± 0.01 (*) | ND |
| MT life time (EB3-YFP) (mean ± SD) | 3.7 ± 0.5 s | 3.5 ± 0.4 s | 5.6 ± 0.6 s (***) | 5.8 ± 0.7 s (***) |
| MT flux (mean ± SEM) | 0.62 ± 0.12 µm/min | 0.55 ± 0.11 µm/min | 0.34 ± 0.13 µm/min (***) | 0.47 ± 0.10 µm/min (***) |
| Parameter\Treatment | Control RNAi | hKID RNAi | KIF4A RNAi | hKID/KIF4A RNAi |
| Mitotic index (MI) (mean ± SEM) | 6.1 ± 0.1% | 7.3 ± 0.3% (*) | 6.7 ± 0.3% | 17.9 ± 1.2% (*) |
| Length of mitosis (median; Q1; Q3) | 35; 30; 40 min | 40; 35; 60 min | 40; 30; 60 min | 90; 65; 150 min (***) |
| Prometaphases (mean ± SEM) | 30.7 ± 3.2% | 42.5 ± 1.6% (*) | 38.4 ± 5.7% | 78.4 ± 3.9% (***) |
| Lagging chromosomes (mean ± SEM) | 3.19 ± 0.52% | 10.77 ± 3.4% | 11.69 ± 2.05% | 25.7 ± 2.25% (*) |
| Cytokinesis failures (mean ± SEM) | 2.37 ± 1.1% | 1.1 ± 0.4% | 4.85 ± 1.8% | 10.45 ± 0.41% |
| ≥3 uncongressed chromosomes | 8.8% | 19.7% | 18.0% | 62.5% |
| Cell death (mean ± SD) | 1.85 ± 0.31% | 2.23 ± 0.26% | 3.06 ± 0.25% (*) | 4.09 ± 0.75% |
| Bipolar spindle size (mean ± SD) | 8.70 ± 0.68 µm | 7.77 ± 0.64 µm (***) | 8.53 ± 0.73 µm | 9.94 ± 1.36 µm (***) |
| Monopolar spindles (mean ± SEM) | 3.98 ± 0.08 µm | 3.12 ± 0.11 µm (***) | 3.78 ± 0.06 µm | 2.85 ± 0.06 µm (***) |
| IK distance (mean ± SD) | 1.01 ± 0.08 µm | 1.00 ± 0.07 µm | 1.27 ± 0.09 µm (***) | 1.24 ± 0.10 µm (***) |
| Oscillation half period (τx) | 52.5 s | 52.5 s | 60 s | 75 s |
| Oscillation speed (mean ± SD) | 1.23 ± 0.21 µm/min | 1.13 ± 0.18 µm/min (**) | 1.13 ± 0.21 µm/min (*) | 1.12 ± 0.17 µm/min (**) |
| Nocodazole sens. MI (mean ± SEM) | 9.9 ± 0.8% (*) | 22.7 ± 1.1% (**) | 4.5 ± 0.3% | 31.9 ± 1.4% (**) |
| MT density (EB1-GFP)(mean ± SEM) | 17.7 ± 0.01 | 14.6 ± 0.03 | 23.5 ± 0.01 (*) | ND |
| MT life time (EB3-YFP) (mean ± SD) | 3.7 ± 0.5 s | 3.5 ± 0.4 s | 5.6 ± 0.6 s (***) | 5.8 ± 0.7 s (***) |
| MT flux (mean ± SEM) | 0.62 ± 0.12 µm/min | 0.55 ± 0.11 µm/min | 0.34 ± 0.13 µm/min (***) | 0.47 ± 0.10 µm/min (***) |
Experiments were performed in HeLa cells except for MT flux and turnover measurements, which were done in U2OS cells. ND, not determined; IK, interkinetochore; sens., sensitivity. *, **, and *** represent Student’s t test P-values of <0.05, <0.01, and <0.001, respectively.