Analysis of centrosome integrity and sister chromatid cohesion in spindle checkpoint–arrested cells
| Treatment . | Separated
chromatids . | Mitotic
index . | Multipolar/
mitotic cellsa . | Poles with a single
centrioleb . |
|---|---|---|---|---|
| Astrin siRNAc | 65.0 ± 2.6 | 16.2 ± 2.0 | 67.3 ± 2.1 | 55.4 ± 13.9 |
| CENP-E siRNA | 9.3 ± 1.2 | 26.3 ± 3.2 | 12.8 ± 3.7 | 16.3 ± 4.8 |
| Nocodazole (16 h)d | 0.3 ± 0.6 | 52.7 ± 2.9 | 23.0 ± 2.0 | 28.7 ± 10.2 |
| Treatment . | Separated
chromatids . | Mitotic
index . | Multipolar/
mitotic cellsa . | Poles with a single
centrioleb . |
|---|---|---|---|---|
| Astrin siRNAc | 65.0 ± 2.6 | 16.2 ± 2.0 | 67.3 ± 2.1 | 55.4 ± 13.9 |
| CENP-E siRNA | 9.3 ± 1.2 | 26.3 ± 3.2 | 12.8 ± 3.7 | 16.3 ± 4.8 |
| Nocodazole (16 h)d | 0.3 ± 0.6 | 52.7 ± 2.9 | 23.0 ± 2.0 | 28.7 ± 10.2 |
Mitotic cells = 100%.
Percentage of poles with a single centriole relative to all poles in multipolar spindles.
Live cell imaging showed that mitotic arrest lasts for ∼10 h before apoptosis occurs.
Cells were synchronized with aphidicolin for 16 h, released for 6 h, and blocked in mitosis for 16 h by adding nocodazole. For the immunofluorescence analysis of mitotically arrested cells, the cells were released into fresh medium for 40 min to allow formation of a mitotic spindle.