Skip to main content
. Author manuscript; available in PMC: 2013 Mar 14.
Published in final edited form as: Antioxid Redox Signal. 2007 Nov;9(11):1803–1813. doi: 10.1089/ars.2007.1579

FIG. 2. The 18-kDa MKK6 fragment binds Rac in 293 cells and RAW cells under conditions that elevate cellular hydrogen peroxide.

FIG. 2

(A, B) The 293 cells were transfected with the following plasmids expressing HA-tagged WT-Rac1, L61-Rac1, N17-Rac1, or WT-Rac1+SOD1. HA-Rac1 was then immunoprecipitated from the 293 cell lysates by using an anti-HA antibody, and the proteins that associated with Rac1 were separated by SDS-PAGE and characterized by Western blotting with the indicated antibodies. In a similar set of experiments, association of endogenous Rac2 with MKK6 was assessed in RAW cells infected with Ad.LacZ or Ad.SOD1. Cell lysate from each condition was immunoprecipitated with anti-Rac2 antibody and evaluated with Western blot. (C) In vitro association of purified GST-CA-MKK6 and GST-WT-MKK6 with His-tagged Rac1. His-tagged Rac1 was immobilized on Dynabeads, and pull-down assays were performed with an equal amount of purified CA or WT GST-MKK6. Proteins bound to Rac1 were evaluated with Western blot by using anti-GST and anti-Rac1 antibodies. All panels are representative for n = 2–5 independent experiments as indicated.