FIG. 7.
Pulse-chase analysis of G1 fusion protein stability. COS7 cells were transfected with ITAM, ΔC51, ITAM-FY, ITAM-YF, and ITAM-FF constructs. In parallel, cells transfected with ITAM constructs were treated with the proteasome inhibitor ALLN for 16 h. Samples were pulsed with [35S]methionine for 1.5 h and were chased for the indicated times (0, 0.5, or 1 h). Lysates of collected samples were immunoprecipitated with Gal4 antibody and were resolved by SDS-PAGE. Samples were visualized by autoradiography, and protein half-lives (T1/2) were calculated by measuring and comparing band densities.