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. 2018 Apr 13;9(28):19597–19612. doi: 10.18632/oncotarget.24696

Figure 3. TRAIL is partially required for the apoptosis induced by CH5126766 and fluvastatin in MDA-MB-231 cells.

Figure 3

(A) The sub-G1 populations after the combined treatment of CH5126766 with fluvastatin in the absence or presence of the pan caspase inhibitor zVAD-fmk. Cells were treated with CH5126766 (40 nM) and fluvastatin (0.3 μM) for 72 h with or without zVAD-fmk (20 μM). DNA contents of the cells were analyzed by flow cytometer. Columns, means of triplicate data; bars, SD; **, P < 0.01. (B) The sub-G1 populations after the combined treatment of CH5126766 with simvastatin in the absence or presence of the pan caspase inhibitor zVAD-fmk. Cells were treated with CH5126766 (20 nM) and simvastatin (0.3 μM) for 72 h with or without zVAD-fmk (20 μM). DNA contents of the cells were analyzed by flow cytometer. Columns, means of triplicate data; bars, SD; **, P < 0.01. (C) The expression of BIM and TRAIL after the combined treatment of CH5126766 with fluvastatin. Cells were treated with CH5126766 (40 nM) and/or fluvastatin (0.3 μM) for 48 h, and the expressions of BIM and TRAIL was analyzed by Western blotting. β-Actin was used as a loading control. (D) The expression of BIM and TRAIL after the combined treatment of CH5126766 with simvastatin. Cells were treated with CH5126766 (20 nM) and/or simvastatin (0.3 μM) for 48 h with or without zVAD-fmk (20 μM). DNA contents of the cells were analyzed by flow cytometer. (E) The knockdown efficacy of siTRAIL validated by Western blotting. β-Actin was used as a loading control. (F) The sub-G1 populations after the combined treatment of CH5126766 with fluvastatin in the TRAIL-depleted cells. Cells were treated with CH5126766 (40 nM) and fluvastatin (0.3 μM) for 72 h after the transfection of each siRNA. DNA contents of the cells were analyzed by flow cytometer. Columns, means of triplicate data; bars, SD; **, P < 0.01.