Procaspase-3 cleavage in cytoplasts.

Cytoplasts were incubated as described in Figure 7. Then whole cytoplast lysates were subjected to SDS-PAGE, and Western blot was performed with an anticaspase-3 polyclonal antibody and an HRP-conjugated secondary antibody. (A) Positive control with procaspase-3 (Jurkat T-cells). (B) Freshly prepared cytoplasts demonstrated only 32-kd procaspase-3. (C) In cytoplasts incubated without stimuli procaspase-3 was cleaved and a 17-kd cleavage product appeared. (D) Addition of G-CSF did not prevent procaspase-3 cleavage and appearance of a 17-kd cleavage product. (E) zVAD-fmk completely inhibited procaspase-3 processing. Results are representative of 3 independent experiments.