Jurkat human acute T-cell leukemia cells were cultured in 96-well plates and treated with the indicated amounts of staurosporine for 4 h. After fixation of cells in the wells, the amount of Cleaved Capase-3 (Asp175) was determined using the R&D Systems Human Cleaved Caspase-3 (Asp175) Cell-Based Infrared ELISA kit (KCBIR835). Signals were normalized to GAPDH staining in each well. Values represent the mean ± the range of duplicate determinations. Analysis of Cleaved Caspase-3 (Asp175) and GAPDH by Western blotting, using the antibodies supplied in this kit, is also shown (inset).
HeLa, HepG2, and HL-60 cells were treated with Staurosporine for 6 h. Treated cells were fixed and analyzed by ICW to measure levels of cleaved PARP-1, using Mitosciences kit MSA43. PARP-1 levels were normalized to cell number. Mean and standard error (n=3) of cleaved PARP-1 are shown. The calculated Staurosporine EC50 values for PARP-1 cleavage are shown. HepG2 cells were resistant to induction of apoptosis under the conditions tested.