Figure 5. hSav1, Mst1/2 and Nek2A regulate centrosome disjunction together with Eg5.
a- RPE-1 cells were treated with the indicated siRNA oligos. Cells were then incubated with nocodazole for 16 h to block MT-dependent centrosome splitting. Prometaphase arrested cells were fixed and stained with γ-tubulin antibodies. Scale bar, 5 μm.
b- Distances between the two centrosomes were analysed from the data in Fig. 5a. Results are from three independent experiments. The S phase distance was measured from cells treated with aphidicolin. n>30 cells counted for each condition. Data are mean ± SEM.
c- RPE-1 cells were treated with the indicated siRNA oligos, enriched in G2 by a single thymidine block/release and subsequently treated with monastrol for 4 h to inhibit Eg5-dependent centrosome splitting. Cells were fixed and stained with α- and γ-tubulin antibodies. Scale bar, 5 μm.
d- Distances between the two centrosomes were analysed from the data in Fig. 5c of cells treated with 100 μM monastrol. Results are from three independent experiments. n >30 cells counted for each condition. Data are mean ± SEM.
e- Cells treated with 50 μM monastrol were analysed for their ability to form bipolar spindles. Results are from three independent experiments. n >30 cells counted for each condition. Data are mean ± SEM.