Effect of CdA treatment on mitochondria in sensitive and resistant cell lines.
(A) Effect of CdA treatment on ΔΨmito. Cells were incubated in the absence (solid line) or presence (dotted line) of 1 μM CdA for 12 hours and then stained with the cationic, lipophilic, membrane potential-sensitive fluorochrome TMRE (25 nM) for 30 minutes. Samples were read on the FL2 channel of a Becton Dickinson FACScan. Data shown are representative of 3 independent experiments. (B) Effect of CdA treatment on release of cytochrome c from mitochondria into cytosol. Cells were incubated in the absence or presence of 100 nM or 1 μM CdA for 12 hours. Cytosols were isolated as described in “Materials and methods,” and the presence of cytochrome c was determined by Western blotting. Jurkat subcellular fractions were included as a positive control to illustrate that although there was no cytochrome c release in resistant cells, the protein was detectable in mitochondrial fractions of untreated Jurkat cells.
