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Figure 5 | Molecular Cancer

Figure 5

From: PTTG/securin activates expression of p53 and modulates its function

Figure 5

Electrophoretic mobility shift assays show the binding of c-myc protein to the c-myc/max sequence. A: Nuclear extract prepared from HEK293 cells transfected either with pcDNA3.1 (lane 1) or pcDNA3.1-PTTG (lane 2) and [32P]-labeled p53 gene promoter sequence carrying a normal c-myc/max binding site. Addition of a 20-fold molar excess of specific unlabeled DNA resulted in almost complete disappearance of the DNA-protein complex (lane 3). An arrow indicates the specific DNA-protein complex. B: Nuclear extracts prepared from HEK293 cells transfected either with pcDNA3.1 (lane 1) or pcDNA3.1-PTTG (lane 2) and [32P]-labeled p53 promoter sequence carrying a normal c-myc/max binding site. Addition of antibody directed against the N-terminal of c-myc resulted in a supershift (lane 3, indicated by an arrow) whereas no supershift was obtained when the [32P]-labeled-D-172/-89 c-myc sequence was used as a probe (lane 5). Addition of antibodies directed against the C-terminal of c-myc did not result in supershift (lanes 4 & 6) when either of the probes was used in the binding reaction indicating that C-terminal of c-myc is not assessable. N.S indicates non-specific complex.

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