Figure 2
From: Inhibition of calpain reduces oxidative stress and attenuates endothelial dysfunction in diabetes
Measurement of ROS production in HUVECs. HUVECs were incubated with normal glucose (NG, 5 mmol/L) or high glucose (HG, 30 mmol/L) in combination with infection with Ad-gal (adenoviral vectors containing beta-gal as a control) or Ad-CAST for 48 h. (A) ROS formation in HUVECs was determined by DCF-DA staining and the nucleus was identified by Hoechst33342 staining. Representative fluorescent pictures of Hoechst33342 (blue signal) and DCF-DA staining (green signal) are shown from 3 different experiments. (B, C) Mitochondrial superoxide flashes were measured in HUVECs. Representative confocal images for the time-course of superoxide flash occurrence from at least 5 cells per experiment in each group. Arrow indicates superoxide flash in a single mitochondrion (B). Quantification of superoxide flashes per μm2 in every 60 seconds (C). Data are given as mean ± SD from 4 different experiments. *P < 0.05 vs. NG treated with vehicle or infected with Ad-gal. #P < 0.05 vs. HG treated with vehicle or infected with Ad-gal.