Figure 3

Functional complementation of ScHOG1 and the multistress behaviour of transformants. The S. cerevisiae wild-type (WT), hog1 and pbs2 strains were transformed with HwHOG1. (A) Ten-fold serially diluted cultures of transformed cells with empty plasmid pRD53 (vector) as control or vector carrying indicated inserts were plated onto YNB-Ura+Gal plates without salt (Control) or supplemented under different conditions (NaCl, KCl, NaCl + pH 8.5, LiCl, sorbitol, pH 8.5), as indicated. Spotted YNB-Ura+Gal plates were also assayed for growth after exposure to UV stress and at low and high temperatures. For the oxidative stress, the cultures were exposed to 50 mM H₂O₂ for 2 minutes prior to plating. (B) Immunodetection with anti-HwHog1p antibodies after SDS-PAGE electrophoresis separation of the denaturated S. cerevisiae cell lysates containing the indicated vectors. Immunodetection with anti-β-actin antibodies served as the sample-loading controls.