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. 2010 Oct 26;60(1):218–226. doi: 10.2337/db10-0021

FIG. 6.

FIG. 6.

The downstream pathway of Src is involved in PI3K/Akt signaling. A: Effects of LY294002 or wortmannin on high-glucose–induced ROS production at 60 min in GK islet cells. After preincubation in the presence of 2.8 mmol/l glucose and 10 μmol/l CM-H2DCFDA for 20 min, dispersed islet cells were incubated in the presence of 16.7 mmol/l glucose with or without 50 μmol/l LY294002 or 0.5 μmol/l wortmannin for 60 min. Fluorescence was represented as fold increases against the value at time zero. Data are expressed as means ± SE (n = 3–5). †P < 0.01. B: Akt phosphorylation in the presence of high glucose with or without exendin-4 or PP2 in GK islets. After preincubation in the presence of 2.8 mmol/l glucose for 30 min, islets were incubated in the presence of 16.7 mmol/l glucose with or without 10 μmol/l forskolin or 10 μmol/l PP2 for 10 min. Islets were lysated and subjected to immunoblot analyses. Blots (50 μg of protein) were probed with anti–phospho-Akt or anti-Akt by stripping and reprobing of the same blots. Intensities of the bands were quantified with densitometric imager. The bar graphs are expressed relative to control value corrected by Akt level (means ± SE). ‡P < 0.001. Representative blot panels of five independent experiments are shown. C: Akt phosphorylation in the presence of high glucose with or without exendin-4 or PP2 in Wistar islets. Representative blot panels of three independent experiments are shown. D: Effects of PD98059 on high-glucose–induced ROS production at 60 min in GK islet cells. Data are expressed as means ± SE (n = 4). E: ERK phosphorylation in the presence of high glucose with or without exendin-4 or PP2 in GK islets. Blots (50 μg of protein) were probed with anti–phospho-ERK or anti-ERK by stripping and reprobing of the same blots. The bar graphs are expressed relative to control value corrected by ERK level (means ± SE). Representative blot panels of three independent experiments are shown. F: Effects of AG1478 on high-glucose–induced ROS production at 60 min in GK islet cells. Data are expressed as means ± SE (n = 5). ‡P < 0.001.