FIG. 6.
Half-life of p53 is increased by IE2-86: pulse-chase. H1299 cells were transfected with expression vectors encoding p53 and the indicated combinations of mdm2, IE1-72, and IE2-86 and pulsed with [35S]Met and then chased with unlabeled amino acids for the indicated times prior to harvesting. (A) Labeled p53 was detected by immunoprecipitation followed by SDS-PAGE separation and autoradiography, and then the resulting film was analyzed with New ImageQuant TL v2003.01 for detecting relative levels of labeled p53 at the indicated chase time points. (B) The amounts of p53, mdm2, IE2-86, and IE1-72 were determined by immunoblotting at the indicated chase time points. “a” refers to transfection with p53, “b” refers to transfection with p53 and mdm2, “c” refers to transfection with p53 and mdm2- IE2-86, and “d” refers to transfection with p53 and mdm2 and IE1-72.