2.3. Dihydroethidium staining

Dihydroethidium staining of the aorta was processed and analyzed as previously described [51]. Briefly, dihydroethidium was prepared at a final concentration of 10 μmol/L in 1X PBS containing 20 % DMSO, applied to 7 μm thick aorta sections, cover-slipped, and incubated for 30 min in a dark, humidified chamber for 30 min. The resulting ethidium fluorescence was viewed using an emission at >580 nm and excitation of 510–550 nm, and fluorescence was quantified on the acquired digital images using Image J fluorescent densitometry (NIH), per unit area. A minimum of 4 tissue sections on 2 slides, n = 3 animals per study group was used to for analyses.