Antisense ON-mediated induction of Bcl-x_Sexpression in human granulocytes leads to apoptosis.

Blood granulocytes were isolated from buffy coats by density centrifugation and treated for 8 hours with DMRIE-C alone (24 μL per 2 × 10⁶ cells) or DMRIE-C combined with AS (or MM) ONs designed to redirect bcl-x pre-mRNA splicing towardbcl-x_S (8 μg per 2 × 10⁶cells). (A) Whole-cell extracts were analyzed by immunoblotting for Bcl-x_L/S and Bcl-2. (B) Apoptosis assays were performed on treated granulocytes using dual-color annexin-V–FITC/PI staining and flow cytometry analyses. The percentage of single- or double-positive cells in the individual quadrants is shown. These results were representative of at least 3 comparable experiments.