Fig 5. NAC deletion relocalizes Ssb to nascent polypeptides and away from prion aggregates.

(A) Fractions from the monosome and polysome peaks of ribosome profile experiments were TCA precipitated prior to SDS-PAGE and Western blotting. More Ssb comigrated with polysomes in the egd1Δegd2Δ strain relative to the WT or to the whole-NAC deletion. Western blots are of the sucrose gradient fractions that contained the monosome and polysome peaks and are representative images from five independent experiments. Quantifications are from five independent experiments, and data are represented as mean ± SEM. See Materials and Methods for full computational details. (B) WT and NAC deletion strains were subjected to co-immunoprecipitation with an anti-Sup35 antibody. Equal amounts of Sup35 were immunoprecipitated across all strains. “Total” and “unbound” fractions were collected and no differences in Ssb protein expression levels were apparent (S3 Fig) Western blots are representative images from three independent experiments. All strains utilized contained [RNQ+] and the strong [PSI+] variant.