The effects of MEK1,2 and Rho-kinase inhibition, as well as AT₁ receptor antagonism, on c-Fos, c-Jun, and Sp1 binding to the MRE. VSMCs were stimulated by Ang II for 1 hour with or without the pretreatment of 50 μM PD98059 (PD), 10 μM Y27632, or 1 μM candesartan (Cand). Nuclear proteins were extracted and incubated with biotinylated MRE prebound to Streptavidin-agarose beads. MRE-associated c-Fos (A), c-Jun (B), and Sp1 (C) was quantitated by Western blot analyses. Representative blots and bar graph quantitations from 3 or more separate experiments are shown. Statistical significance with P < .001 by ANOVA is marked as ^**.