Figure 5.

In vitro phosphorylation of Nrf2 by PKC. In vitro kinase assays were performed by using purified Nrf2 and catalytic subunits of rat brain PKC. (A) Nrf2 (2 mM) was incubated with 5 mM PKC and 2 mCi of [γ-³³P]ATP for the indicated times at 30°C. The reaction products were analyzed by SDS/PAGE and autoradiography. [γ-³³P]ATP incorporation was quantitated by a phosphor imager. (B) Identical kinase assays were carried out for 30 min at 30°C in the presence of various concentrations of staurosporine or Ro-32–0432.