Figure 5
From: Inhibition of Bcl-2/Bcl-xL and c-MET causes synthetic lethality in model systems of glioblastoma
C-MET and Bcl-2/Bcl-xL inhibition suppresses protein synthesis and stabilize pro-apoptotic Noxa. (A) LN229 and U87 GBM cells were treated with ABT263, Crizotinib or the combination of both (24 h) (concentrations in μM). At the end of the treatment period, cells were incubated for 2 h with puromycin. Thereafter, whole cell protein lysates were harvested and analyzed by capillary electrophoresis for the expression of puromycin (protein synthesis activity in cancer cells after various treatments referred to as SUNSET assay). Cycloheximide (CHX) serves as a positive control (concentrations are in μg/ml) ABT: ABT263, Criz: Crizotinib. (B) LN229 or U87 GBM cells were exposed to cycloheximide in the presence or absence of the combination treatment of ABT263 and Crizotinib. Thereafter, whole protein lysates were isolated and analyzed by capillary electrophoresis. ABT + Crizo: ABT263 1 μM + Crizotinib 2 μM. Noxa-S and Noxa-L refer to the exposure times. L: long exposure; S:short exposure. (C) LN229, U87 and stem like GBM cells, NCH644, were treated as indicated for 24 h. RT-PCR was performed for the expression of Mcl-1 and Noxa. Ctrl: Control, ABT: 1 μM ABT263, Crizo: 2 μM Crizotinib.
