Figure 4.
Figure 4. Adaphostin induces cell death through generation of ROS. (A) ROS generation. Example of a mean fluorescence histogram for CLL cells. CLL lymphocytes were incubated for 30 minutes in diluent (open black line), 30 μM adaphostin (open gray line), or 30 μM adaphostin added about 2 hours after 20 mM NAC (black line with gray shading). (B) NAC attenuates adaphostin-induced cell death. CLL B cells were cultured in the presence or absence of NAC (20 mM) or the broad-spectrum caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp fluoromethylketone (z-VAD-fmk) (50 μM). After 2 hours, adaphostin (10 M) was added to some cultures. CLL B cells were recovered from cultures after 24 hours, and the percentage of viable cells was determined by annexin/PI staining using flow cytometry analysis (mean ± SD; n=6). Y error bars indicate standard deviation.

Adaphostin induces cell death through generation of ROS. (A) ROS generation. Example of a mean fluorescence histogram for CLL cells. CLL lymphocytes were incubated for 30 minutes in diluent (open black line), 30 μM adaphostin (open gray line), or 30 μM adaphostin added about 2 hours after 20 mM NAC (black line with gray shading). (B) NAC attenuates adaphostin-induced cell death. CLL B cells were cultured in the presence or absence of NAC (20 mM) or the broad-spectrum caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp fluoromethylketone (z-VAD-fmk) (50 μM). After 2 hours, adaphostin (10 M) was added to some cultures. CLL B cells were recovered from cultures after 24 hours, and the percentage of viable cells was determined by annexin/PI staining using flow cytometry analysis (mean ± SD; n=6). Y error bars indicate standard deviation.

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