Figure 8.

Activation of p53 by pTpT. Cultures of H1299 cells were transfected with either control vector or the p53 expression vector, treated with pTpT and processed for the electromobility shift assay as described in Materials and Methods. Lane 1, extract from cells transfected with the p53 expression vector. Lane 2, cells were transfected with the p53 expression vector and incubated with 100 μM pTpT. Lane 3, same as lane 2 but the binding reaction contained 0.1 μg mAb421. Lane 4, same as lane 3 but with a 100-fold excess (10 pmol) unlabeled wild-type p53 consensus sequence. Lane 5, same as lane 3 but with a 100-fold express (10 pmol) unlabeled mutant p53 consensus sequence. The consensus sequence DNA/p53 complex (C.S. DNA/p53) and the supershifted complex (C.S. DNA/p53/Ab421) are indicated by arrows.