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Figure 5 | Molecular Cancer

Figure 5

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 5

API-1 induces GSK3-dependent Mcl-1 phosphorylation and degradation. A, The given cell lines were treated with 5 μM API-1 for different times as indicated. B, The given cell lines were pre-treated with 10 μM SB216763 for 30 min and then co-treated with 5 μM API-1 for an additional 2 h. C, The indicated cell lines were transfected with control (Ctrl) or GSK3α/β siRNA and 48 h later were exposed to DMSO or 5 μM API-1 for additional 4 h. D, H1299 cells were pre-treated with 10 μM SB216763 (SB) for 30 min and then co-treated with 5 μM API-1 for additional 4 h. The cells were then washed with PBS 3 times, refed with fresh medium containing 10 μg/ml CHX and cultured for the indicated times. E, The indicated plasmids were transfected into 293 T cells. After 48 h, the cells were treated with 5 μM API-1 for an additional 4 h. After the aforementioned 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. The results were plotted as the relative Mcl-1 levels compared to those at the time 0 of CHX treatment (D). LE, longer exposure.

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