Centrosome clustering was impaired in p53-/- MEF cells with extra centrosomes.

Cells were treated with Cyto-B (0.5 µg/ml) for 12 h, then released for 8 h, and stained with antibodies against γ-tubulin and α-tubulin for centrosomes and microtubules, respectively. Nuclei were stained with DAPI. Note that only cells with more than 3 centrosomes at metaphase were analyzed. The percentage of cells with indicated mechanism was shown separately. Mean ± s.d. from three independent experiments.*p2 test, compared with MEF p53+/+ cells.

将细胞以0.5 μg/ml的细胞松弛素B（Cyto-B）处理12小时，随后洗脱并恢复培养8小时；分别使用靶向中心体的抗γ微管蛋白（γ-tubulin）抗体，以及靶向微管的抗α微管蛋白（α-tubulin）抗体进行免疫荧光染色。细胞核采用4',6-二脒基-2-苯基吲哚（DAPI）染色。注：本实验仅分析分裂中期中心体数量超过3个的细胞。分别统计并展示了对应目标机制的细胞占比。数据以三次独立实验的平均值±标准差（standard deviation, s.d.）呈现。采用*p2检验，与p53野生型小鼠胚胎成纤维细胞（mouse embryonic fibroblast, MEF）进行比较。