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. Author manuscript; available in PMC: 2016 May 18.
Published in final edited form as: Leukemia. 2015 Aug 12;30(1):219–228. doi: 10.1038/leu.2015.222

Figure 7. BIM knockdown can rescue AUY922-induced apoptosis in KOPT-K1 T-ALL cell line.

Figure 7

(a) BIM was silenced by lentiviral shRNA knockdown in KOPT-K1 cells and protein expression was assessed by western blotting. Two independent shRNAs (shBIM #3 and #4) were compared with a control shRNA targeting luciferase (shLuc). (b) KOPT-K1 cells transduced with each of BIM-targeting shRNAs (shBIM #3 and #4) or a control shLuc were treated with graded concentrations of AUY922 for 48 h. Cell viability values are shown as mean ± s.d. percentages of the untreated control value in triplicate experiments. (c) KOPT-K1 cells transduced with each of BIM-targeting shRNAs (shBIM #3 and #4) or a control shLuc were treated with DMSO or 30 nM of AUY922 for 36 h. These cells were fixed and assessed for apoptosis and cell-cycle distribution by flow cytometric analysis following TUNEL/PI double labeling. Values represent mean ± s.d. percentages of TUNEL-positive cells in triplicate experiments. ***, P < 0.001 by two-sample, two-tailed t test. The representative dot plots images in this assay are shown in Supplementary Figure 8. (d) KOPT-K1 cells transduced with each of BIM-targeting shRNAs (shBIM #3 and #4) or a control shLuc were treated with 30 nM of AUY922 or DMSO (AUY922 0 nM) for 36 h. Whole-cell extracts were analyzed by immunoblotting with each specific antibody.