50l of 2�� reaction buffer con taining 10 mM DTT was added to each sample . 50M final concentration substrates this explanation for caspase 3 and caspase 9 were added to each sample for a total volume of 100l and incubated for 180 m at 37 C. Free pNA released from the labeled synthetic substrate on cleavage by active caspase was measured on a fluorescence plate reader at 405 nm. SCID/Human Xenograft Female Inhibitors,Modulators,Libraries ICR SCID mice were obtained from Taconic Labo ratories and were housed and treated in the Wayne State University School of Medicine under an approved protocol. Four week old mice were injected intraperitoneally with 5 106 WSU FSCCL cells. ApoG2 was injected 25 mg/kg QD 5 days either IP or intravenously. Mice were observed Inhibitors,Modulators,Libraries daily and eutha nized when they appeared ill. Animals activity, weight and survival were monitored three times a week.

mice were sacrificed when they developed hind region paraly sis, had decreased activity Inhibitors,Modulators,Libraries and weight loss of 15% or more, or death was felt to be imminent. Necropsy was carried Inhibitors,Modulators,Libraries out and the extent of macroscopic disease was identified with all major organs being taken for microscopic patho logical examination. Major organs included the brain, femur, heart, kidney, liver, lungs, pan creas, retroperitoneal fat, and spleen. Peripheral blood smears were examined for evidence of circulating lym phoma cells. Survival curves were created using the product limit of Kaplan and Meier, and compared using the log rank test. The end point for assessing anti lymphoma activity was calculated by percent increase in host life span. %ILS 100 MDD /MDD of the tumor bearing control mice.

Statistical analysis Apoptosis induction by AnnexinV/PI stains and AO/EB were compared to control by the student t test. Survival functions were estimated using the Kaplan Meier method and compared by the log rank test. P values 0. 05 were Inhibitors,Modulators,Libraries considered statistically significant. All statistical analyses were evaluated using GraphPad Prism 4 Results Effect of ApoG2 on WSU FSCCL Cells The structure of ApoG2 is shown in figure 1A. To study if ApoG2 is effective in our FL cell line, WSU FSCCL, we determined the baseline expression levels of anti apop totic, Bcl 2, Bcl XL and Mcl 1 and expression levels of pro apoptotic, Bax, Bak, and Bad proteins. Our west ern blots show that our FL cell line has high expression of anti apoptotic proteins and pro apoptotic protein Bax, but low expression of pro apop totic proteins.

This profile predicts that ApoG2 should be an vitamin d effective agent in this model. To study cytotoxic effects of ApoG2, WSU FSCCL cells were exposed to increasing concentrations of the SMI for 24 to 72 h. We exposed WSU FSCCL cells to ApoG2 at concen trations of M. ApoG2 significantly inhibited the growth of WSU FSCCL in a concentration and time dependent manner. For example, ApoG2 at a concentration of 10. 00M ApoG2 inhibited the growth of WSU FSCCL cells by 90% at all incubation times.