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Supplementary figures
Figure S1.SAC-dependent sensitization of MDA-MB231 cells to death ligands
(A) Representative images of morphology of cells treated with different anti-mitotic drugs in presence or absence of reversine. MDA-MB231 cells were treated with nocodazole (250 ng/ml), taxol (1 M), BI2536 (100 nM) or STLC (20 M) in presence or absence of reversine (1 M) for 24 hours. (B)MDA-MB231 cells were incubated for 15 hours in complete medium with or without nocodazole (250 ng/ml), taxol (1 M), BI2536 (100 nM) or STLC (20 M), in presence or absence of reversine (1 M). After this incubation, cells were treated for 8 hours with TNF(100 ng/mL, upper panel) or FasL (100 ng/mL, lower panel) and apoptosis was determined.Error bars represent S.D. from three independent experiments. *P<0.05,**P<0.01, *** P<0.001.
Figure S2.Sensitization to TRAIL-induced apoptosis by Cdc20 depletion.
(A) HeLa cells were transfected and synchronized as described in legend to figure 2A during 24 hours. Then, cells were released in fresh medium and after 15 hours, TRAIL (50 ng/mL) was added for 6 hours. (B) Hela cells were transfected and synchronized as in (A). After 24 hours, cells were released in presence or absence of reversine. Simultaneously, control cells were also treated or not with nocodazole. After 15 hours, TRAIL (50 ng/mL) was added for 6 hours. In both cases, apoptosis was measured by flow cytometry as the percentage of cells with sub-G1 DNA content. (C) Sensitization to TRAIL-induced apoptosis by over-expression of non-degradable cyclin B. U2OS-CycB-TR cells were synchronized in G1/S by thymidine block. Afterwards, expression of non-degradable cyclin B was induced in some cultures by addition of doxycycline (1 g/mg). After 16 hours, TRAIL (250 ng/mL) was added for 7 hours. Apoptosis was measured as the percentage of cells with sub-G1 DNA content. Error bars represent S.D. from three independent experiments. **P<0.01, *** P<0.001.