Figure 4.

Ste6–13p, Ste6–90p, and Ste6–166p define two classes based on aberrant metabolic stability. The metabolic stability of ER-localized ste6 mutants was examined by pulse-chase analysis. Cells were pulse-labeled for 10 min with Express ³⁵S, and the label was chased for the indicated times (min). Wild-type and mutant forms of Ste6-HAp were immunoprecipitated from total extracts with the anti-HA antibody, and immunoprecipitates were analyzed by SDS-PAGE on 8% gels. Bands were detected and quantified by PhosphorImager analysis using Imagequant software. The Ste6p half-life (t_1/2) is indicated at the bottom of each panel. Strains were the same as in Figure 1.