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. 2015 Oct 23;7(6):6576–6592. doi: 10.18632/oncotarget.5878

Figure 6. Pro-apoptotic BH3-only protein BIM is up-regulated by MLN0128; knockdown of Bim significantly rescues MLN0128-mediated MCC cell death.

Figure 6

A. Increased Bim expression upon MLN0128 treatment by Western blotting. MCC cells were treated with MLN0128 for 24 hours at indicated concentrations. Tubulin was used as a loading control. B. Up-regulated Bim expressions upon MLN0128 treatment by qRT-PCR. MCC cells were treated with MLN0128 for 24 hours at the indicated concentrations. Samples were run in triplicate and normalized to MRPS2 mRNA to determine relative expression (means ± SEM). *p < 0.05 compared with controls. C. Decreased Bim expression in MCC cells transduced with Bim shRNA. A non-targeting scramble shRNA served as a control. Tubulin was used as a loading control. D. Decreased cell death in Bim-knockdown MCC cells upon MLN0128 treatment. Bim shRNA or non-targeting control shRNA transduced MCC cells were treated with MLN0128 at indicated concentrations, followed by Annexin-V and PI staining and flow cytometry analysis. E. A significant reduction in MLN0128-mediated cell death in MCC cells transduced with Bim shRNA compared with non-targeting control shRNA (*p < 0.05). Data are presented as the mean ± SEM of triplicate experiments.