FIG. 4.
Stability of WT and T125D mutant TAgs. R2-JCVE, R2-T125A, and R2-T125D cells were pulsed with [35S]methionine-containing medium for 1 h. Cell lysates were collected at regular intervals (at 0, 3, 6, 12, 24, and 36 h postlabeling), immunoprecipitated with PAb 962 antibody, and electrophoresed in SDS-15% polyacrylamide gels. TAg bands were detected by autoradiography, and band intensities were quantitated using a phosphorimager and ImageQuant 5.2 software. The percent radiolabeled protein relative to that measured at 3 h postlabeling (100%) was plotted against time. The T125A mutant TAg was not detected, whereas WT and T125D mutant TAgs exhibited similar stabilities. Results from one of three independent experiments are shown.