Figure 9.

Sis1 chaperone module is required for Sis1 to coimmunoprecipitate with Rnq1. (A) Coimmunoprecipitation of Rnq1 with chimeric Hsp40s. Extracts from sis1Δ-expressing Sis1, SYS, and YSY were prepared under nondenaturing conditions and Sis1 was immunoprecipitated from cell extracts. The presence of Rnq1 in the immunoprecipitates was then detected by Western blot with α-Rnq1 sera. The amount of Rnq1 in total cell extracts was also probed by Western blot, and these data are presented in the bottom panel. (B) Relative quantities of Sis1, SYS, and YSY that were immunoprecipitated from cell extracts. Western blots of immunoprecipitates generated for A were reprobed with α-Sis1 sera to detect the quantity of Sis1, SYS, and YSY that was immunoprecipitated. The bottom panel represents a Western blot of total cell extract probed with α-Sis1 sera. P.I. denotes control immunoprecipitations carried out with Sis1 preimmune (P.I.) sera. ^* IgG denotes the position of IgG in the sera that was used for the immunoprecipitations that is present on membranes and cross reacts with the second antibody utilized to develop these Western blots.