FIG. 9.

Effects of skn7-dock2 and skn7-T437A mutations on the oxidant (ox)-dependent phosphorylation of Skn7. Shown is the oxidant-dependent phosphorylation of (A) wild-type SKN7 (as seen in Fig. 2), (B) skn7-dock2, and (C) skn7-T437A. TCA extracts prepared from JF1904 (skn7Δ) cultures carrying SKN7 expression plasmids (SKN7⁺, pXH1853; skn7-dock2, pXH1941; and skn7-T437A, pXH1854) and left untreated (−) or treated (+) with oxidant (2 mM H₂O₂ for 20 min) were subjected to SDS-10% PAGE for 2.5 h at 200 V. Lambda phosphatase (Ph) was added where indicated to 40 U (C, lanes 2 and 3) or 100 U (A and C, lanes 4 and 5) for 30 min at 30°C according to the manufacturer's recommendations. (A and C) Phosphatase inhibitors (In) were added to all lanes except for those labeled Ph. A dashed white line is added to the figure based on the migration of untreated wild-type control samples on the outside edges of each gel.