FIG. 8.

In vivo Nox1 gene silencing blocked CCN2(IV)-induced vascular responses. C57BL/6 mice received a single i.p. injection of recombinant CCN2(IV) (2.5 ng/g body weight) or vehicle and were sacrificed after 24 h. Some mice were pretreated with a Nox1 morpholino (10 mg/kg, retroorbital injection) 24 h before CCN2(IV) administration. (A) Nox1 protein expression was evaluated by Western blot. (B) O₂^•− production was determined by an increase in 2-OH-E⁺ generation by HPLC analysis of DHE fluorescence. (C) Protein nitrosylation and phosphorylated p65 NF-κB subunit (p-p65) levels were evaluated by immunostaining in paraffin-embedded aortic sections. (D) Levels of phosphorylated IκBα protein were evaluated in aortic protein extracts by Western blot. (E) Gene expression was evaluated by real-time PCR in aortic samples. Data are expressed as mean±SEM of fold increase over saline of 8–10 animals per group. *p<0.05 versus saline. ^†p<0.05 versus CCN2(IV). To see this illustration in color, the reader is referred to the web version of this article at www.liebertpub.com/ars