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. 2015 Sep 5;6(29):27580–27595. doi: 10.18632/oncotarget.4768

Figure 5. Induction of SIRT1 protein degradation by CPZ.

Figure 5

A. HCT116 cells were treated with CPZ in a dose- and time-dependent manner, and SIRT1 protein and mRNA levels were analyzed using Western blotting and RT-PCR assays, respectively. SIRT1 protein expression was quantified using ImageJ software. B. HCT116 cells were treated with cycloheximide (CHX, 50 μg/mL) in the presence or absence of CPZ (10 μM) for 0–12 h, and the SIRT1 protein level was analyzed using Western blotting. C. HCT116 cells were treated with CPZ (0, 10, and 20 μM) in the presence or absence of MG132 (25 μM), and SIRT1 protein expression was analyzed using Western blotting. D. HCT116 cells were pretreated with SP600125 (10 and 20 μM) and then treated with CPZ for 24 h. Protein lysates were immunoprecipitated with an antiSIRT1 antibody and blotted with antiubiquitin (upper panel) and anti-SIRT1 (lower panel) antibodies. E. HCT116 cells were pretreated with SP600125 (20 μM) and then incubated with CPZ (10 and 20 μM) for 24 h, and the SIRT1 protein level was assessed using Western blotting and quantified using ImageJ software.