FIG. 2.

Individual p53 mutants demonstrate variable levels of p53 specific transactivation and apoptosis. (A) Mutant and wild-type p53 expression vectors were cotransfected into H1299 cells with luciferase constructs containing (p21p) or lacking (p21pΔ1.1) p53-responsive elements. PCR amplification of a known mutant p53 species (143-2M) containing a C170W mutation in exon 5 was performed in parallel. Background transactivation of constructs lacking p53 response elements was set to a value of 1. (B) Mutant and wild-type p53 expression vectors and RFP-expressing plasmids were cotransfected into SAOS-2 cells by calcium phosphate precipitation (7). At 36 h posttransfection, apoptosis was measured by labeling the ends of fragmented DNA with FITC. RFP-positive cells were gated the percentage of cells dual positive for RFP and FITC are shown.