Fig. 4.

Hsp82 and Hsc82 mutants exhibit defective Ssa1 interaction in vivo. Pull-down assays from yeast cell lysates were carried out as described in Materials and Methods using lysates from cells expressing wild-type or mutant His–Hsc82 (A and B) or wild-type or mutant His–Hsp82 (C and D). (A and C) The proteins from the pull down and the cell lysate were analyzed by SDS/PAGE and visualized by Coomassie staining and immunoblot analysis; shown are representatives of at least three independent experiments. (B and D) Quantification of Ssa1 bound to wild-type or mutant His–Hsc82 (B) or to wild-type or mutant His–Hsp82 (D). The ratio of Ssa1 bound to each of the His–Hsc82 or His–Hsp82 mutants relative to wild type is presented as the mean ± SEM. *P < 0.05, **P < 0.01, n.s., not significant (ANOVA; see Materials and Methods).