Effect of Bcl-XLL down-regulation on cell survival in K-299 cells. K-299 cells were transiently transfected with a pEGFP control vector (left panels) or ASBcl-XL-GFP plasmid (right panels) encoding full-length Bcl-XL antisense cDNA. Cultures containing at least 60% of transfected cells were used for further analysis by Western blot and apoptosis determination. (A) Western blot analysis of total lysates for the expression of Bcl-XL was performed at the indicated time points after transfection. NPM/ALK levels are presented as a protein loading control. (B) At 80 hours after transfection, cultures were treated for 16 hours with DMSO (middle panels), 10 μM etoposide (right panels), or left untreated (left panels). Apoptosis induction was assessed using an annexin V binding assay and flow cytometric analysis.
