Activation of caspases by TNFα. Neutrophils were incubated in the presence of cycloheximide (10 μg/mL), alone (control), or with TNFα (10 ng/mL) or GM-CSF (50 U/mL) in the presence or absence MG132 (50 μM) and Z-VAD-FMK (50 μM). After 5-hour incubation, samples were removed and levels of caspases-3, -8, -9, and -10 (or GAPDH as a loading control) were detected by Western blotting. Typical result of 3 separate experiments.