Figure 7.

Bartter mutant degradation is mediated by the Hsp70 chaperone in yeast. A–E, cycloheximide chase reactions were performed, and lysates were blotted with anti-HA antibody to measure the stability of ROMK and the indicated mutant proteins over 90 min. The degradation of ROMK was measured in SSA1 (open squares) or ssa1-45 (closed squares) mutant yeast, which were incubated at 37 °C for 20 min before the start of the chase, and the incubation was continued at 37 °C. Data represent means of six independent experiments ± S.E. (error bars). In each experiment, a representative Western blot is shown at the bottom. G6PD was used as a loading control. *, p < 0.05; **, p < 0.01; ***, p < 0.001.