FIG. 11.

Effect of YAP1 overexpression on oxidant (ox) sensitivity and the phosphorylation of oxidative stress-defective skn7 mutants. (A) Cultures of skn7 mutants with and without a high-copy YAP1 expression plasmid were spotted (10-fold dilutions starting at 10⁶cells/ml) on plates lacking or containing oxidant. Yeast strains included JF1904 (skn7Δ) carrying the following SKN7 expression plasmids: SKN7⁺, pXH1853; skn7-T437A, pXH1854; skn7-T449A, pXH1939; skn7-dock2, pXH1941; and the high-copy (hc) YAP1 expression plasmid (pSEY18-YAP1). (B) Effect of the high-copy YAP1 expression plasmid on the migration of the skn7-dock2 mutant protein. TCA extracts were prepared from JF1904 (skn7Δ) carrying the skn7-dock2 expression plasmid (pXH1941) and the high-copy YAP1 expression plasmid (pSEY18-YAP1) or vector. Extracts were prepared from cultures treated with 2 mM H₂O₂ (20 min) (+) or left untreated (−) and were subjected to SDS-10% PAGE for 2.5 h at 200 V. A dashed white line is added to the figure based on the migration of two untreated wild-type samples run as controls on the outside edges of the gel.