Fig 4. As₄O₆ induces MMP (ΔΨm) depolarization, activates of caspases and cleavage of PARP, and DR5 as well as regulation of Bcl-2, IAP family.

The cells were incubated at 0, 0.1, 0.5, 1, 2 and 5 μM concentrations of As₄O₆ for 24 h. (a) The cells were stained with JC-1 and incubated at 37°C for 30 min. The mean JC-1 fluorescence intensity was assessed by a flow cytometer. As₄O₆ induced depolarization of MMP (ΔΨm) at 2 and 5 μM concentrations. (b) Total cell lysates were resolved by SDS-polyacrylamide gels and transferred onto nitrocellulose membranes. The membranes were probed with the anti-caspase-3, anti-caspase-8, anti-caspase-9 and anti-PARP antibodies. (c) The membranes were probed with the antibodies against DR4, DR5, Bcl-2 and IAP family members. To confirm equal loading, the blot was stripped of the bound antibody and reprobed with the anti ß-actin antibody. The proteins were visualized using an ECL detection system. The data are shown of three independent experiments.