FIG. 2.

Increased expression and phosphorylation of p47^phox and ROS generation in the ZDF rat islets compared with the ZLC islets ROS. Levels were measured in isolated islets from ZLC and ZDF rats after incubation with DCHFDA (10 μmol/L) for 30 min. Islets were washed with ice-cold phosphate-buffered saline and sonicated. An equal amount of protein was used to quantitate 2′,7′-dichlorofluorescin fluorescence. A: Data are expressed as percent control and are mean ± SEM (error bars) from islets from four rats in each group. In a separate experiment, islets from the ZLC or the ZDF rats were lysed using radioimmunoprecipitation assay buffer. Equal amounts of lysate proteins were resolved by SDS-PAGE. Expression of phosphorylated and total p47^phox was determined by Western blotting. A representative blot is provided for total p47^phox (B) and phospho-p47^phox (D). Densitometric quantitation of total p47^phox (C) and phosphorylated p47^phox (E). Data are mean ± SEM (error bars) from islets from four rats in each group. *P < 0.05 vs. ZLC rat islets.