Bcl-2-ASO–induced down-regulation of Bcl-2 correlates with apoptosis and precedes apoptosis in low Bcl-2–expressing ARP-1 myeloma cells.
ARP-1 cells (low Bcl-2) were seeded at 5 × 105/mL in T-25 flasks in FCS medium. Bcl-2-ASO was added at 5 μg/mL, and cultures were harvested following 0, 1, 2, 3, 4, and 5 days of treatment. Bcl-2-ASO was replenished every other day. Apoptosis (A) was determined by the annexin V method and Bcl-2 protein by Western immunoblotting (B). The quantitation for Bcl-2 was done by densitometry (C). Loading controls for Bcl-2 protein were performed by prerunning of gels and staining with Coomassie blue as detailed in “Materials and methods.” Other experimental details are as in Figure 1. Representative results from 2 experiments are shown. Note that down-regulation of Bcl-2 precedes apoptosis for every time point tested.
