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. 2014 Aug 21;42(16):10433–10447. doi: 10.1093/nar/gku763

Figure 2.

Figure 2.

Both NOXA and BIM contribute to MYC-dependent apoptosis. (A) DanG cells were siRNA transfected as indicated. Sixty hours after the transfection, cells were treated with bortezomib for additional 12 h. Western blot of MYC, BIM and NOXA. Same extracts were transferred to two membranes and both were controlled by β-actin for equal loading. (B) DanG cells were treated as in (A). NOXA, BIM and MYC mRNA expression was determined by qPCR using PPIA mRNA as reference. Student's t-test *P < 0.05, n = 3. (C) 3T9 MYClox/lox-CreER mouse embryonic fibroblasts cells were treated for 3 days with 4-hydroxytamoxifen (TAM; 250 nM) and subsequently treated with bortezomib for additional 12 h or left as vehicle-treated controls. Western blot analysis of MYC and BIM expression (β-actin: loading control). (D) Stable transfected sh control and shBIM DanG cells were transiently transfected with a control siRNA or a NOXA siRNA. After 60 h, cells were treated with bortezomib for additional 12 h. Western blot of BIM, NOXA and β-actin (loading control). (E) Relative caspase 3/7 activity of stable transduced sh control and shBIM DanG cells, which were transiently transfected with a control siRNA or a NOXA siRNA for 48 h. Caspase 3/7 activity was measured 24 h after bortezomib treatment. One-way ANOVA *P < 0.05, n = 3.