Figure 3.

Analysis of H1299 cell lines stably transfected with mutant p53 plasmids. Clonal cell lines were generated by stable transfection with the plasmid pRC-CMV containing p53 cDNAs encoding the wild-type p53 protein (lanes 1–5), the transactivation mutant, 22-23 (lanes 6–10), the DNA binding mutant, R175H (lanes 11–15), the proline deletion mutant, ΔproAE (lanes 16–20), or the temperature-sensitive murine DNA binding mutant, A135V (lanes 21 and 22). Cell lines were analyzed for p53 protein expression by Western blot analysis of p53-specific immunocomplexes. Cell lysates were immunoprecipitated with the panspecific antibody, pAb421, which recognizes both mutant and wild-type conformations of p53 (lanes 1–21) or the negative control antibody, pAb419 (lane 22). Immunocomplexes were resolved by 10% SDS/PAGE, transferred to an Immobilon-P membrane (Millipore), and immunoblotted with pAb421. The positions of full-length p53 and the ΔproAE proteins are indicated.