FIG. 5.

Hog1p phosphorylation with various SLN1 truncation mutations. Antiphosphotyrosine immunoblot analysis was performed on immunoprecipitated Hog1p separated by SDS-PAGE. (A) Rich medium; (B) defined medium. Odd-numbered lanes, low-osmolarity medium; even-numbered lanes, high-osmolarity medium. Lanes 1 and 2, full-length SLN1 with restriction sites (Fig. 1A); lanes 3 and 4, ΔTMD1 construction (Fig. 1C); lanes 5 and 6, KD construction (Fig. 1E). Duplicate anti-Hog1p immunoblots demonstrated that the same amounts of Hog1p were immunoprecipitated in all cases.