Figure 6.

Duration of Hog1 nuclear accumulation is reduced in cells lacking Msn2 and Msn4 transcription factors. Logarithmically growing strain W303-1A (WT) and YM24 (msn2,4Δ) transformed with pVR65–WT (HOG1–GFP) were examined by fluorescence microscopy under iso-osmotic (selective medium) or hyperosmotic growth conditions (5 min after addition of NaCl into selective medium to a final concentration of 0.4 M) at the indicated time points (A) or by determining the phosphorylation profile of Hog1–GFP (B). Samples were taken at the time points indicated, and protein extracts were prepared. Western blots were analyzed with anti-active p38 antibody (asterisks indicate the positions of proteins that are nonspecifically recognized by antibody) or with anti-GFP antibody (bottom panel) to determine the protein level of Hog1–GFP.