Figure 2.

4EGI-1-induced apoptosis involved the intrinsic pathway. (A) Primary myeloma cells, HMCLs and PBL were treated with the given concentration of 4EGI-1 for 18 h in culture conditions. Cell death was then assessed by AnnexinV staining. (B) Myeloma cell lines were treated with 50 μℳ 4EGI-1 for 18 h in culture conditions. Equivalent amounts of cell lysates were separated by SDS–PAGE and then immunoblotted with PARP-1, anti-caspase-3 and anti-caspase-9 antibodies. Actin was used as a loading control. (C) Cells were pre-incubated with 20 μℳ caspase inhibitors (z-VAD-fmk or caspase-9 inhibitor) for 4 h before to be treated by 4EGI-1 for 18 h. Cell death was expressed as the percentage of cells with subG1 DNA content.