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. 2014 Oct 6;9(10):e108839. doi: 10.1371/journal.pone.0108839

Figure 5. CpdA increases cellular levels of polyubiquitinated proteins and sensitizes cells to b-AP15 induced cell death.

Figure 5

A. HCT116 cells were pre-treated with or without (control) 10 µM cpdA for 16 hours, exposed to the indicated concentration of b-AP15 for 1 hour, then incubated for a further 16 hours in drug-free medium. Lysates were subjected to immunoblotting for K48-linked polyubiquitin, PARP and β-actin (loading control). B. HCT116 cells were treated as above. Following treatment, the number of dead cells was measured by Trypan-blue staining. Bar chart shows mean +/− SD of three independent experiments. Statistical significance was calculated using the Student's t-test. P values * = 0.05 and ** = 0.01. C. and D. hTERT-RPE1 cells were treated and analyzed as above. E. and F. Human diploid fibroblasts were treated and analyzed as above, and the number of dead cells was determined 36 hours after drug treatment.