Fig. 1. Analysis of the amount of nonsense suppression and soluble Sup35 in [PSI⁺] variants. (A) Five-fold serial dilutions and growth of [PSI⁺] variants on rich media (YPD, 30°C, 4 days) and media lacking adenine (SC–Ade, 30°C, 14 days). (B) Quantitation of translational read through of [PSI⁺] variants. (C) Dot-blot analysis of Sup35 solubility in [PSI⁺] variants. Proteins from the top fraction of each sucrose cushion were normalized, serially diluted in 2-fold increments and applied to a PVDF membrane. Top, reactivity with a Sup35-specific antibody (Patino et al., 1996). Bottom, membrane stained with a non-specific protein stain, Ponceau-S. The amount of total protein per column is indicated at the top.