Figure 3.

BV6/Dexa cotreatment stimulates mitochondrial perturbations prior to cell death. (a) Cells were treated for indicated time points 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 three independent experiments performed in triplicate are shown. (b and c) Cells were treated for indicated time points 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. MMP was assessed by TMRM staining and flow cytometry. Mean and S.D. of three independent experiments performed in triplicate are shown; *P<0.05; **P<0.01. (d and e) Cells were treated 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 after indicated time points (d) or MitoSOX staining after 4 h (e) and flow cytometry. Mean and S.D. of three independent experiments performed in triplicate are shown; *P<0.05. (f) 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. Respiration was determined by the Oxygraph system (basal respiration and respiration after addition of indicated concentrations of oligomycin or FCCP). Mean and S.D. of three independent experiments are shown