FIGURE 1.

Establishing AA-induced p53 mutants in platform MEFs by IMCE. A, strategy for generating sets of p53 mutants in a common genetic background with the p53 platform/IMCE procedure. B and C, platform MEFs transfected with TOP plasmids containing the indicated p53 mutations together with PhiC31 integrase plasmid were selected for 3 days with puromycin before growth for a further 3 days in the absence of antibiotic selection. The ability of various TOP constructs containing the indicated p53 mutations to produce colonies after 7 days in antibiotic selection was determined and expressed as colonies formed per 1 × 10⁴ cells seeded for transfection. Colonies were fixed and stained with crystal violet (CV) before extraction with 10% acetic acid. Absorbance was read at 590 nm (arbitrary units; AU) and taken as a measure of both colony number and size.