FIG. 1.

Dopaminergic cells contain NADPH oxidase components and 6-OHDA leads to Nox1induction and Rac1 activation. (A) mRNA of Nox isoforms and subunits were detected using RT-PCR in N27 cells treated with 6-OHDA (100 μM) for 6 h. GAPDH, internal control; control, vehicle treated. (B) 24 h 6-OHDA treatment increased Nox1 expression in a dose-dependent manner as examined by immunoblot analysis. Signal intensity was measured by Quantity One software and shown as a fold increase. β-actin, internal control. (C) GTP-bound active Rac1 was increased by 6-OHDA treatment for various durations (1 h and 6 h) as determined by the active GTPase pull-down assay. Nox1, coprecipitated with active Rac1, was detected in the same blot using Western blot analysis. Total Rac1 and β-actin were demonstrated as internal controls. Images are representative of three independent experiments.