Figure S1 - Hsp110 Chaperones Regulate Prion Formation and Propagation in S. cerevisiae by Two Discrete Activities

Deletion of SSE1 impairs [PSI+] propagation. (A) Tetrad analysis of [PSI+] sse1�� cross with [PSI+] wild-type cells. Wild-type cells of the weak ([PSI+]w) or strong ([PSI+]s) phenotypes were crossed with the corresponding [PSI+] sse1�� cells. Diploids were sporulated and [PSI+] phenotypes of segregants were assessed by growth on rich media (YPD) after 2 days and media lacking adenine (-Ade) after 3�C6 days. Upper panels: [PSI+]w tetrad. Lower panels: [PSI+]s tetrad. (B) Centrifugation analysis of Sup35 and Rnq1. Aggregation of Sup35 and Rnq1 in [PSI+]w wild-type and sse1�� cells was assessed by centrifugation analysis and Western Blotting. (C) Agarose gel electrophoresis of Sup35 monomers and polymers. Monomeric and polymer forms of Sup35 in different strains were analyzed using SDD-AGE (semi-denaturing detergent-agarose gel electrophoresis) and Western Blotting. (D) Chaperone protein levels in sse1�� cells or cells overexpressing Sse1. Levels of the chaperones Hsp104, Ssa1 and Ssb1 were determined by Western Blotting using G6PDH as a loading control. (5.86 MB EPS)