Figure 3.

hMOF interacts with Nrf2. (A) hMOF can interact with Nrf2. Plasmids expressing HA-Nrf2 or Flag-hMOF were transfected or co-transfected into 293T-cells. 48 h after transfection, total proteins were subjected to IP and IB analysis with indicated antibodies. (B) H₂O₂ promotes the interaction between hMOF and Nrf2. 293T-cells were transfected with plasmids expressing HA-Nrf2 or Flag-hMOF. Thirty-six hours later, the cells were left untreated or were treated with 100 μM H₂O₂ for additional 12 or 24 h. Cell lysates were subjected to IP and IB analysis with the indicated antibodies. (C) GST pull-down assay showing Nrf2 binds directly to hMOF. In vitro Nrf2 bound purified recombinant GST-hMOF. GST served as the pull-down control. Equal amounts of input Nrf2 were measured by CBB staining, while the pull-down products were detected with anti-Nrf2 antibody.