FIG 3.

CgHog1 phosphorylates CgRds2 at 1.5 M NaCl. (A) Immunoprecipitation of CgRds2-Myc was performed in the wild-type (wt) and Cghog1Δ strains at 0 M NaCl and 1.5 M NaCl, followed by Western blotting using anti-Myc antibody. The arrow indicates the phosphorylation band of CgRds2. (B) Extracts prepared from CgRds2-Myc-expressing wild-type cells, grown at 0 M NaCl and 1.5 M NaCl, were treated with alkaline phosphatase and phosphatase inhibitor as indicated. The arrows indicate the phosphorylation band of CgRds2. (C) Immunoprecipitation of phosphorylated CgRds2 was performed in the wild-type (wt), Cghog1Δ, Cgrds2^4A, and Cgrds2^2A strains at 0 M NaCl and 1.5 M NaCl, followed by Western blot analysis using anti-phosphoserine/threonine antibody. Quantification of relative phosphorylation levels of CgRds2 in the wild-type (wt), Cghog1Δ (D), Cgrds2^4A (E), and Cgrds2^2A (F) strains at 0 M NaCl and 1.5 M NaCl. β-Actin was used as a loading control. All data are presented as mean values of three independent experiments. Error bars indicate the standard deviations. ***, P < 0.001.