Fig 2.

Kinetics and coincidence of Rim13 and Rim20 localization. Log-phase cells expressing Rim13-GFP (A) or Rim20-GFP (B) grown in YPD were shifted to CSM plus 0.1 M HEPES adjusted to pH 8.3 and then imaged at the indicated times after pH shift on three independent days. The number of cells displaying fluorescent foci after the shift was quantified and plotted. (C to F) A RIM20-GFP vps4Δ strain transformed with pRS315 plasmid bearing RIM13-tdTomato was imaged in CSM for colocalization of GFP and tdTomato fluorescence. For a representative field of cells, the signals from GFP (C), tdTomato (D), and both GFP and tdTomato (E) are shown. Scale bar, 5 μm. The rectangles in panels C and D illustrate regions that were scanned for signal intensity. (F) Signal intensities of GFP (green line) and tdTomato (red line) across select regions are graphed. The four panels display the scans across four fields, and the left-hand panel is the scan of the region indicated by the rectangles in panels C and D.