FIG. 7.

CCN2(IV) activates NF-κB via Nox1 in VSMCs. (A) Time course of phosphorylation of p65 NF-κB subunit (p-p65) in mice VSMCs treated with 50 ng/ml CCN2(IV). (B) Concentration–response curve [CCN2(IV) 1–100 ng/ml; 60 min]. (C, D) VSMCs were transfected with p22phox, Nox1 siRNA, or scramble siRNA before CCN2(IV) stimulation for 60 min. Figure shows a representative Western blot, and the quantification of p-p65 protein levels expressed as mean±SEM of fold-change over control of eight (A), four (B), five (C), and three (D) independent experiments. *p<0.05 versus control. ^†p<0.05 versus CCN2(IV). (E) Cells were preincubated 60 min with apocynin (0.3 mM) or ML-171 (1 μM) or (F) transfected with p22phox before CCN2(IV) stimulation for 60 min. Localization of p65 NF-κB subunit was evaluated by confocal microscopy. (E, F) A representative image of three independent experiments. siRNA, small interfering RNA molecule. To see this illustration in color, the reader is referred to the web version of this article at www.liebertpub.com/ars