FIG. 2.

Degradation of wt Cot and trunc-Cot in HEK293 transfected cells. Immunoprecipitation of ³⁵S-labeled wt Cot and trunc-Cot from HEK293 cells transfected with 10 μg of pcINEO HA-wt Cot or 10 μg of pcINEO HA-trunc-Cot after a 20-min pulse-label and a chase for the times indicated (20, 40, 60, 120, and 240 min). The proteins immunoprecipitated were resolved by SDS-PAGE and visualized by autoradiography. The radioactivity incorporated was quantified using an Instant Imager. The figure shows one experiment, and the graph shows the mean ± standard deviation of the radioactivity incorporated in four experiments. The radioactivity incorporated into wt Cot or trunc-Cot after the 20-min pulse was considered 100%. The degradation of both proteins correlated with one-phase exponential equations fit to each data set, with R² = 0.975 for wt Cot and R² = 0.9736 for trunc-Cot. The graph also shows a representation of the decay as a log plot. ▴, wt Cot; ▪, trunc-Cot.