Figure 3

Cleavage of caspase 9, 3 and PARP protein and induction of Caspase 3, 9 activity and resulting DNA fragmentation in TW-37 treated lymphoid cell lines. A) WSU-FSCCL cells were exposed to TW-37 (0 to 750 nM) at 24, 48 and 72 hr. Caspase 3, 9, 8 and PARP protein cleavage was detected by Western blot (3A.1). Or cells were treated with TW-37 at 0, 250 and 500 nM on white 96-well plate for 4, 8, 12 and 24 hr. Caspase 3, 9 activities were determined by Luminescence immediately on 96-well plate (3A.2). WSU-FSCCL cells treated with TW-37 at 250 and 500 nM. DNA fragmentation was seen evident after 48 hr (3A.3). (3B1.) Caspase 9 and 3 enzyme activation by TW-37 was also determined using WSU-DLCL2 lymphoid cell line. Cells exposed to 300, 400 nM of TW-37 for 4 or 8 hr, caspase 9 and 3 activity was detected. (3B.2) WSU-DLCL2 cells treated with TW-37 at 500 and 750 nM, DNA fragmentation was evident after 48 hr.