FIG. 3.

Capsaicin-induced ROS generation involves NOX3 activation. (A) UB/OC-1 cells exposed to capsaicin demonstrated increased ROS generation, as measured by H₂DCFDA fluorescence. UB/OC-1 cells were pretreated with scramble or NOX3 siRNA for 48 h, the culture media were replaced with fresh media, and cultures were treated with vehicle or capsaicin (2.5 μM). Capsaicin increased ROS was abrogated by NOX3 siRNA. The transfection of cells with NOX3 siRNA alone did not alter H₂DCFDA fluorescence. (B) Cells pretreated with vehicle or BAPTA-AM (10 μM) for 30 min before capsaicin administration showed reduced capsaicin-mediated ROS generation. The addition of BAPTA-AM alone was not associated with any appreciable change in ROS generation. Scale bar (lower right panel) represents 10 μm. Data presented in (A) and (B) were replicated at least three times with similar results. BAPTA-AM, 1,2-Bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid tetrakis (acetoxymethyl ester). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article at www.liebertonline.com/ars).