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Figure 7 | Molecular Cancer

Figure 7

From: The E3 ubiquitin ligases β-TrCP and FBXW7 cooperatively mediates GSK3-dependent Mcl-1 degradation induced by the Akt inhibitor API-1, resulting in apoptosis

Figure 7

Both the SCF E3 ligases, β-TrCP and FBXW7, are involved in mediating API-1-induced Mcl-1 degradation. A, H1299 cells were transfected with control (Ctrl) or FBXW7 siRNA and 48 h later were exposed to DMSO or 5 μM API-1 for an additional 4 h. Moreover, WT and FBXW7-KO HCT116 cell lines were treated with 5 μM for 4 h. B, H1299 cells were pre-treated with the indicated concentrations of MLN4924 and then co-treated with the given concentrations of API-1 for an additional 4 h. C and D, H1299 cells were transfected with the given siRNAs for 48 h and then exposed to 5 μM API-1 for another 4 h. After these treatments, the cells were harvested for preparation of whole-cell protein lysates and subsequent Western blotting. Protein levels were quantified with NIH Image J software and were normalized to actin. E, H1299 cells were transfected with the given siRNAs and after 48 h were harvested for extraction of total RNA. RT-PCR was then conducted to detect the expression of the indicated mRNAs.

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