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. 2013 Nov 13;8(11):e78766. doi: 10.1371/journal.pone.0078766

Figure 5. 280B down-regulates p53 protein by inducing Mdm2 expression in prostate cancer cells.

Figure 5

(A, B, E) C81 cells were transfected with (A, E) control siRNA (−) or 280B siRNA (+) or (B, E) infected with Empty adenovirus (−) and 280B adenovirus (+) and subjected to real-time RT-PCR and Western blotting to measure the expression of Mdm2 and 280B. (C, D) C81cells were transfected with 0.1 µg Mdm2-Luc and co-transfected with (C) Control siRNA (−) and 280B siRNA (+), (D) Empty pCIneo (−) and 0.2 µg and 0.4 µg 280B, and monitored for 280B activity by measuring Luciferase activity. (E) C81cells were co-transfected with empty pCIneo(−) or Mdm2 (+) and control siRNA (−) or Mdm2 siRNA (+), followed by Western blotting to measure the cytosolic (C) or nuclear (N) levels of p53, 280B, and Mdm2. For all experiments, β-actin served as a loading control. β-tubulin and RARa were used as markers for cytosolic and nuclear fractions, respectively (E). All p53 protein levels were relative to the first condition, and it was set to 1. Asterisks * and ** indicate statistical significances of P<0.05 and P<0.01, respectively.