Effect of PI 3-K inhibitors on apoptosis in HEC 1-A, RL 95-2 and Ishikawa cells. Control (■), LY294002 (○) and Wortmannin (▲). 2 × 106 cells were plated for 0, 6, 12, 24 h and cultured in the presence of medium and LY294402 or Wortmannin. Cells were trypsinized, pooled with floating cells and collected for Hoechst nuclear staining (right panel) or TUNEL analysis (left panel) to count apoptotic cells. Data represent the mean ± SEM of 4 independent experiments.