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. 2015 Dec 4;7(3):2809–2822. doi: 10.18632/oncotarget.6465

Figure 6. Effect of Ulocuplumab (BMS-936564)-mediated apoptosis in CLL cells is caspase-independent.

Figure 6

A. Cells were treated for 6 hrs with either 200 nM of Ulocuplumab (BMS-936564) or isotype control (IC). 100 μg of protein was taken from each treatment for caspase analysis. Caspase 2, 3, 8 and 9 were activated upon treatment with the Ulocuplumab (BMS-936564) antibody. There was a significant difference between normal versus CLL cells for caspase 2, 3, 8, and 9. Caspase activity of cells incubated in media only was used as a baseline control. Each value is expressed as mean ± S.D. of two independent experiments. B. CLL cells were incubated with Ulocuplumab (BMS-936564) (200 nM), F-ara-A (10 μM), and Etoposide (30 μM) for 48 hrs either alone or in combination with different concentrations of a pan-caspase inhibitor, Z-VAD-FMK (10, 30, 90 μM). CLL cells viability was analyzed by CD19+/CD5+/Annexin-V staining followed by flow cytometry. Statistical significance was determined by using Dunnett's multiple comparison test (* p < 0.05; ** p<0.01, *** p < 0.001). Z-VAD inhibited apoptosis in a dose dependent manner when apoptosis was induced by chemotherapy controls but not by Ulocuplumab (BMS-936564) (untreated media control vs. 0-90 μM Z-VAD, p < 0.0001).