FIG. 1.

FLIP inhibits H/R-induced apoptosis in MLEC associated with caspase 3 activation and cytochrome c release. MLEC at 30% confluence were cultured in serum-free media in the presence of 10⁶ CPU/ml of adeno-FLIP or adeno-lacZ for 3 h and then restored to normal medium. Two days later, the cells were exposed to hypoxia (95% N₂, 5% CO₂) for 24 h and then restored to normoxic culture conditions (95% air, 5% CO₂). At the indicated reoxygenation times, total lysates or cytosolic fractions were prepared as described in Materials and Methods. The fractions were subjected to immunoblotting (IB) to detect caspase 3 or cytochrome c.