Fig. 5.

BAF and zVAD-FMK block caspase activity, even after 24 hr of camptothecin exposure. E18 cortical neurons were treated with camptothecin alone or in combination with flavopiridol (Flavo; 1 μm), BAF (100 μm), or zVAD-FMK (VAD; 100 μm). Ten or 24 hr later, soluble (A) or particulate (B) neuronal lysates (50 μg) were generated as described in Materials and Methods and subjected to SDS-PAGE on a 12% (A) or 10% (B) gel. After Western immunoblotting with the anti-N-Nedd antibody (B) (1:250) or the C-2-10 monoclonal anti-PARP antibody (1:5000) (B), the bands were visualized by ECL (Amersham). The arrows indicate the 37 kDa cleavage product for caspase-2 (A) and the 85 kDa PARP cleavage product (B).