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. 2018 May 9;8:7373. doi: 10.1038/s41598-018-25802-0

Figure 3.

Figure 3

Protein levels of anti- and pro-apoptotic Bcl-2 family members upon treatment with c-MET inhibitor, ABT263 or the combination of both. (A) U87, NCH644 and LN229 GBM cells were treated with increasing concentrations of Crizotinib (in μM) for 1d. Thereafter, whole cell lysates were prepared and analyzed by capillary electrophoresis on the Wes Instrument (Proteinsimple Inc.) for the expression of Mcl-1, Bcl-2, Bcl-xL, Noxa and Vinculin. (B) LN229 and U87 GBM cells were treated with ABT263 (ABT), Crizo (Crizotinib) or the combination for 1 d (concentrations in μM). Whole cell protein lysates were prepared and analyzed for the same markers as in A. (C) LN229 and U87 GBM cells were transfected with two siRNAs targeting Mcl-1. Thereafter, cells were treated with either ABT263, Crizotinib or the combination (1d). And then harvested, fixed, stained with propidium iodide and analyzed by flow cytometry. Shown are means and SD. (D) LN229 and U87 GBM cells were transfected with Noxa or Bak specific siRNAs and treated with the combination of 1 μM ABT263 (ABT) and 2 μM Crizotinib (Crizo) for 1d. Cells were processed and analyzed as in C. Shown are means and SD. (E) LN229 and U87 GBM cells were transfected with Bak, Noxa or Mcl-1 specific siRNAs. Thereafter, whole protein lysates were prepared and analyzed for the expression of Noxa, Bak and Mcl-1 by capillary electrophoresis on the Wes Instrument (Proteinsimple, Inc.). Concentrations are in μM.