Ficoll gradient-purified mononuclear cells from PB from newly diagnosed B-lineage ALL, from healthy volunteers, CML CD34+ cells, or BV173 or SUPB15 cells were cultured at 1 × 106 cells/ml in the presence of increasing concentrations of ABT-199 (0, 0.001, 0.01, 0.1, 1 µM) for 3 h followed by staining with 50 nM TMRE (tetramethylrhodamine ethyl ester) for 20 min at 37 °C. Separately, cells were treated with the ionophore FCCP (5 µM) (Carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone) as a positive control. MOMP of viable cells was determined by flow cytometry.
Pre-treatment of PDX cells co-cultured on bone marrow derived mesenchymal stem cells (MSCs) or of BV173 cells with dexamethasone (100 or 50 nM) and dasatinib (50 or 0.5 nM) was performed for 48 h prior to TMRE assay.
Cell culture, cloning of lentiviral constructs, apoptosis analysis, proliferation assays, immunoprecipitation, immunoblotting and statistical methods are described in detail inSupplementary Materials .
Pre-treatment of PDX cells co-cultured on bone marrow derived mesenchymal stem cells (MSCs) or of BV173 cells with dexamethasone (100 or 50 nM) and dasatinib (50 or 0.5 nM) was performed for 48 h prior to TMRE assay.
Cell culture, cloning of lentiviral constructs, apoptosis analysis, proliferation assays, immunoprecipitation, immunoblotting and statistical methods are described in detail in
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