Mitochondrial damage by nucleosides in B-CLL cells.
(A) Mitochondria from B-CLL cells were nitrogen isolated by cavitation, treated for 10 minutes with the indicated concentrations of 2CdATP (▪), CaFdATP (■), F-Ara-ATP (●), and dATP (○), stained with 40 nmol/L DiOC6 for 10 minutes at room temperature, and rapidly analyzed by flow cytometry. (B) B-CLL cells were treated with 10 μmol/L of the nucleosides for 8 hours, then permeabilized with 0.03% of digitonin and fixed with 4% paraformaldehyde. AIF release was detected by staining with specific antibody and with fluorescence-labeled secondary Alexa 488 and measured by flow cytometry. (C) Human leukemia T-lymphoblastoid CEM cell lines, transfected with bcl-2–expressing vector (CEM/bcl-2) (○) or control vector (CEM/NEO) (●), were treated with 1 μmol/L of the nucleosides. Membrane potential analyses and caspase analysis were performed at the indicated time points. Error bars represent the SD obtained from 3 independent experiments.
