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Supplementary Data shLuc sh14-3-3 sh14-3-3 179 shLuc sh14-3-3 sh14-3-3 179 Asyn Noco 2.1% 6.1% 2.4% 16.5%11.4%1.1% 5.9%3.9%2.5% 52.5%10.0%4.9% shLuc shBubR1 2N 4N>4N 2N 4N>4N 2N 4N>4N 2N 4N>4N 2N 4N >4N 2N 4N>4N Figure S2 Supplementary data Figure S1 and S2. HeLa cells were transfected with the indicated shRNAs, and dead cells were removed by washing 24 hrs later. Thirty-six hrs after the transfection, the cells were treated with nocodazole (660 nM) for another 36 hrs, fixed in 70% ethanol, stained with propidium iodide, and analyzed by flow cytometry to determine DNA content. (S1) The dead and dying cell population (R1) and the live cell population (R2) are indicated, and the percentages of each population are shown. (S2) The aneuploid populations (>4N) are indicated, and their percentages are shown. Asyn, asynchronized; Noco, nocodazole treatment. R1 R2 R1 R2 R1 R2 shLucshBubR1shMad2 sh14-3-3 Figure S1 shLucshBubR1shMad2 shLuc R1 R2 R1 R2 R1 R2 R1: 42.4% R2: 50.2% R1: 26.9% R2: 66.1% R1: 44.7% R2: 47.4% R1: 88.2% R2: 5.3% R1: 88.7% R2: 5.1% R1: 88.2% R2: 5.2% Noco (36h) Noco (36h) FSC SSC FSC SSC
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Supplementary data Figure S3. HeLa cells were transfected with the indicated shRNAs or Bcl2 expressing vector and dead cells were removed by washing 24 hrs later. 36 hrs after the transfection, the cells were treated with nocodazole for another 24 hrs with or without Z-VAD-FMK and then spread and incubated for 12~14 days to allow colony formation. Colonies were stained with crystal violet and counted. The surviving colonies (%) was normalized by setting the percentage of surviving colonies in the control wells to 100. Error bars represent standard deviation. Supplementary Data Figure S3 0 50 100 0 50 100 0 50 100 shLuc sh14-3-3 179sh14-3-3 616 VehicleNocodazole VehicleNocodazole VehicleNocodazole + Z-VAD-FMK w/ Mock vectorw/ Bcl2 expressing vector w/ or w/o Z-VAD-FMK Surviving Colonies (%)
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Supplementary Data Supplementary data Figure S4. HeLa cells were transfected with the indicated shRNAs, and dead cells were removed by washing 24 hrs later. Thirty-six hrs after the transfection, the cells were treated with nocodazole (660 nM) for another 36 hrs, Cisplatin (24uM) and Rapamycin (20nM) for another 48 hrs. Relative viability was assessed by MTS assay. Data are expressed by the mean of percent cell viability compared to control. Error bars represent standard deviation. Figure S4 RapamycinNocodazoleCisplatin Luc 14-3-3 179 sh14-3-3 616 shRNA Luc 14-3-3 179 sh14-3-3 616 Luc 14-3-3 179 sh14-3-3 616 50 100 Relative viability (%)
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