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Fig. 3. Reversible change in the localization of GFP-Rat8p. Yeast cells in exponential phase were treated with 10% ethanol for 30 minutes, collected, and transferred to fresh SD medium without ethanol. GFP-Rat8p fluorescence was visualized before ethanol treatment (w/o stress) after ethanol treatment for 30 minutes (10% EtOH) and 5 minutes after the shift to fresh SD medium. Cells at each stage were also fixed for in situ hybridization using Cy3-labeled SSA4 probes to detect poly(A)+ RNA and DAPI staining.