Fig. 1
Induction of mitochondrial apoptosis in BCR-ABL + ALL by BH3-mimetics. a (Left) BV173 were treated with various concentrations of ABT-199, ABT-737, and WEHI-539 for 3 h followed by staining with TMRE to determine MOMP of viable cells by flow cytometry. Treatment with the ionophore FCCP served as positive control, and fluorescence of untreated cells was set as 100%. (Right) PI staining to monitor cell viability was performed by flow cytometry after additional 21 h. Values are expressed as means ± SD (n = 3). b PB mononuclear cells from newly diagnosed BCR-ABL-negative ALL (n = 13), BCR-ABL-positive ALL (n = 7), CML CD34+ cells (n = 7) and PB mononuclear cells from healthy volunteers (n = 6) were incubated with 1 µM ABT-199 for 3 h for MOMP induction followed by TMRE staining and FACS analysis of viable cells. p-value was calculated using Student’s t-test (*p < 0.05). c BV173 cells were treated with 100 nM ABT-199 for the indicated periods of time. Cellular lysates were immunoprecipitated with anti-BCL2, anti-BCLXL, and anti-MCL1 antibodies, respectively. Total cellular lysates and immunoprecipitates were subjected to western blot analysis using the indicated antibodies. Quantification of band intensities was normalized to the amount of precipitated target protein. d BV173 cells were stably transduced with lentiviral constructs encoding specific shRNA targeting BIM or a control sequence and treated with various concentrations of ABT-199, dexamethasone, imatinib or doxorubicin for 48 h. PI staining was performed to monitor cell viability. Values are expressed as means ± SD (n = 3). p-values were calculated using Student’s t-test (**p < 0.01, ***p < 0.001)
