Figure 3
DCZ3301 induces MMP decreases and enhances caspase activation. (a) OCI-LY8 cells treated with DCZ3301 (4, 8, 12 and 16 μM) and (b) NU-DUL-1 cells treated with DCZ3301 (5, 10, 15 and 20 μM) for 24 and 48 h were stained with JC-1 to examine the level of MMP by flow cytometry. (c) Assessment of OCI-LY8 and NU-DUL-1 cells treated with DCZ3301 by JC-1 showed change in MMP. Columns show the loss of MMP represented as means±S.D. (*P<0.05). (d) Western blotting analysis was performed to explore the protein levels of caspase family, including cleaved caspase-3, capase-8 and caspase-9, PARP and B-cell lymphoma (Bcl) family, such as Bcl-2, Bcl-xL and Bax. (e) NU-DUL-1 cells incubated with or without pan-caspase inhibitor Z-VAD-FMK were both treated with 15 μM DCZ3301 for 48 h, followed by Annexin-V/propidium iodide (PI) analysis on flow cytometry. (f) Columns show the percentage of Annexin-V positive cells from three independent experiments, which are shown as means±S.D. (*P<0.05). (g) Normal PBMCs from three healthy volunteers treated with different concentrations of DCZ3301 (0, 20 and 40 μM) for 48 h were stained with Annexin-V/PI and analyzed by flow cytometry; the ratio of apoptotic cells shown as mean±S.D.
