Cleavage of caspase-8 and caspase-3 in PMNs.

Whole-cell lysates from freshly isolated PMNs (lane 1), from PMNs cultured for 6 hours without additions (lane 2), with 150 μM zVAD-fmk (lane 3), with 20 ng/mL TNF-α (lane 4), or with the TNF-α/zVAD-fmk combination (lane 5) were subjected to SDS-PAGE. Western blot was performed with anti–caspase-8 monoclonal Abs (A). Then the blot was reprobed with anti–caspase-3 polyclonal Abs (B). The expression of Bax protein determined by anti-Bax polyclonal Abs was used as a measurement for equal protein loading (C). Results are representative of 3 independent experiments.