FIGURE 1.

Sensitive oxidation of PRL by H₂O₂. A, COS7 cells were transfected with the indicated expression constructs and then treated with the indicated concentrations of H₂O₂ for 10 min. Cell lysates were subjected to nonreducing or conventional (reducing) SDS-PAGE followed by immunoblot analysis with anti-Myc antibody. Area between arrows, oxidized proteins containing intermolecular disulfide bonds; arrowhead, reduced proteins; asterisk, oxidized proteins containing intramolecular disulfide bonds. B, HEK293 cells were treated with the indicated concentrations of H₂O₂ for 10 min, and cell lysates were subjected to anti-PRL immunoblot analysis. C, HEK293 cells were transfected with the indicated constructs (without any tags) and then treated with 500 μm H₂O₂ for 10 min. Cell lysates were subjected to anti-PRL immunoblot analysis. D, COS7 cells were transfected with the indicated constructs and then treated with 500 μm H₂O₂ for 10 min. Cell lysates were subjected to anti-Myc immunoblot analysis. E, COS7 cells were transfected with Myc-PRL3 (upper panels) or Myc-PTEN (lower panels) and then treated with the indicated concentrations of H₂O₂ for 10 min. Cell lysates were subjected to immunoblot analysis with the indicated antibodies. F, HEK293 cells were treated with the indicated concentrations of H₂O₂ for 10 min, and cell lysates were subjected to immunoblot analysis with the indicated antibodies.