Simulation of shapes of Sup35NM-GFP oligomers in [PSI⁺] lysates. (A) SDD-AGE of yeast lysates obtained from [PSI⁺] cells treated with (right) or without (left) GuHCl. An anti-Sup35 antibody was used for Western blotting detection. Sup35m represents the monomeric form of Sup35NM-GFP. (B) Representative normalized FAFs of GFP prepared from lysates of [PSI⁺] cells (black), Sup35NM-GFP from [psi⁻] cells (blue), Sup35NM-GFP from [PSI⁺] cells (green) without GuHCl treatment, and Sup35NM-GFP from [PSI⁺] cells with GuHCl treatment (red) are shown. Solid lines show the FAF fitting curves. (C) Relation between diffusion constants and molecular masses (mm). Oblique lines indicate simulated plots for Perrin’s factors (F = 1–5). Mean diffusion constants were calculated from three independent FCS experiments. The molecular masses of GFP (black) and Sup35NM-GFP (blue) were calculated from their amino acid sequences. A molecular mass of 4,200 kD was used as the mean for the Sup35NM-GFP oligomers (green), which had a mean diffusion constant of 4.6 ± 2 µm²/s, calculated from the corresponding FCS data. The diffusion constant of Sup35NM-GFP in the [PSI⁺] lysate with GuHCl treatment was categorized into two groups because the FAFs fit well to a two-component model. Molecular masses of 8,000 kD (red) and 20,000 kD (orange) were used as representatives for the two different diffusion constants of 4.7 ± 3 µm²/s and 2.8 ± 1 µm²/s, respectively.