Figure 2

YM155 induced apoptosis in SK-NEP-1 cells. YM155 induced apoptosis, DNA fragmentation and activation of caspase 3 in SK-NEP-1 cells. (A) Annexin V analysis showed the cells treated with YM155 50nM and 100nM for 6 and 12 hours, much more cells showed apoptotic feature compared with control group. These analyses were repeated three times, and the apoptotic cells in 6 hours was YM155 50nM: 5.9% ±2.3%,YM155 100nM 42.4% ±8.9% compared to DMSO group: 1.5%±0.4%; apoptotic cells in 12 hours was YM155 50nM: 31.5% ±5.7%, YM155 100nM 45.1% ±11.3% compared to DMSO group 6.5% ±2.1%. (B) Cell cycle analysis the cells treated with YM155 after 12, 24 and 36 hours. As expected, DNA fragmentation was observed after 12 hours treatment and increased in a time dependent manner. These analyses were repeated three times, and the apoptotic cells in YM155 50nM group: 12 hours 8.7% ±1.4%, 24 hours 23.2% ±6.7%, 36 hours 74.0%±9.2%; apoptotic cells in YM155 100nM group: 12 hours 18.5% ±3.7% , 24 hours 25.4% ±9.2% , 36 hours 60.9% ±14.2%. (C) Western-blot analysis the activation of caspase 3 in cells treated with YM155. After 6 and 12 hours treatment with 10nM, 50nM, 100nM and 200nM YM155, cleaved caspase-3 was observed.