Figure 1.

Formation of fluorescent structures begins as an early focus and develops into line and dot structures with distinctive growth patterns. (a) Sup35PrD-GFP was overexpressed in [psi ⁻][PIN ⁺] strains for approximately 18 hours. Cells were categorized as having diffuse cytoplasmic fluorescence (diffuse), the presence of a small focus/several foci on a diffuse background (early foci; indicated with arrow), rings/lines, or dots. The percentage of cells containing each fluorescent structure was determined by counting more than 700 cells from 3 independent transformants. Standard error is shown. (b) After 18 hours of overexpression, diffuse cells were visualized for an additional 12 hours using 3D time-lapse microscopy. Early foci (left top panel, arrow), mid (middle panel), and late stages (right panel) of ring formation (time shown as HH:MM:SS) relative to the first appearance of early focus are shown. The lower panel shows a more detailed time-lapse, with different colored arrows (white and pink) indicating different ends. All images shown are maximum projection. (c) Z-stacks (indicated as μm), corresponding to specific time points (rows), is shown for a single cell forming a single focus over time. 3.5 μm is expanded for more detail. The diameter of the growing dot structure was measured, as indicated by representative line. Arrows indicate when the focus is visible in each z-stack.