Figure 4.

BV6/Dexa-induced loss of MMP and cell death depend on Bak. (a) Cells were treated for 6 h with BV6 (Tanoue: 3 μM BV6; Jurkat: 5 μM BV6) and/or 200 μM Dexa, Jurkat cells were additionally treated with 20 μM zVAD.fmk. Bak activation was determined by IP using an active conformation-specific antibody. Protein expression of Bak was analyzed by western blotting. β-Actin served as loading control. (b) ALL cells were transiently transfected with two distinct siRNAs targeting Bak or control siRNA. Protein expression of Bak was analyzed by western blotting. β-Actin served as loading control. (c) Cells were treated for 24 h with BV6 (Tanoue: 3 μM BV6; Jurkat: 5 μM BV6) and/or 200 μM Dexa, Jurkat cells were additionally treated with 20 μM zVAD.fmk. Cell death was determined by FSC/SSC analysis and flow cytometry. Mean and S.D. of at least three independent experiments performed in triplicate are shown; *P<0.05. (d) Cells were treated for 6 h with BV6 (Tanoue: 3 μM BV6; Jurkat: 5 μM BV6) and 200 μM Dexa, Jurkat cells were additionally treated with 20 μM zVAD.fmk. Loss of MMP was determined by TMRM staining and flow cytometry. Mean and S.D. of three independent experiments performed in triplicate are shown; *P<0.05. (e) Cells were treated for 4 h with BV6 (Tanoue: 3 μM BV6; Jurkat: 5 μM BV6) and/or 200 μM Dexa, Jurkat cells were additionally treated with 20 μM zVAD.fmk. ROS production was determined by CellROX staining and flow cytometry. Mean and S.D. of three independent experiments performed in triplicate are shown; NS, not significant