Ectopic overexpression of Smac/DIABLO sensitizes Jurkat cells to Epo B– or Apo-2L/TRAIL–induced apoptosis.

Jurkat cells were transiently transfected with either a control vector (pcDNA3.1 Zeo) or a vector containing full coding sequence of Smac/DIABLO (pcDNA3.1 Smac). Forty-eight hours later, cells were harvested and Smac/DIABLO expression was determined by immunoblot analysis of the cell lysates by using anti-Flag and anti-Smac antibodies (A). Twenty-four hours after transfection, the cells were treated with 10 nM Epo B (B) or 2.5 ng/mL Apo-2L/TRAIL (C) for 24 hours. Following this, the percentage of apoptotic cells was determined by annexin V staining followed by flow cytometry. Values represent the mean ± SE of 3 independent experiments.