FIGURE 9.

Hsp31 and Sup35 are not mutually co-localized, and Hsp104 and Hsp70 are not altered. A, most cells with high levels of expression of Hsp31 and Sup35 exhibited diffuse cytoplasmic localization for Hsp31 that overlapped with Sup35 (top). Occlusion of Hsp31 from the Sup35 aggregate was also observed, as evidenced by the decreased DsRed signal at aggregate sites (white arrows, bottom panel). B, SDS-resistant Sup35 aggregates are suppressed by Hsp31. Cellular lysates were subjected to SDD-AGE and SDS-PAGE. Overexpression of Sup35-PrD-EYFP initiated the formation of Sup35 aggregates, which were detected with anti-GFP antibody. C, Hsp31 overexpression does not alter the expression levels of Hsp104 and Hsp70. Cells of the BY4741 strain harboring the pGPD-HSP31-DsRed or vector control (pGPD) were grown overnight, and samples were prepared as described in the legend to Fig. 3. Immunoblots (IB) were probed with anti-Hsp104, anti-Hsp70, anti-DsRed, and anti-actin antibodies.