Fig. 8.

Afobazole elevates lymphatic endothelial cell nitric oxide (NO) production. A: trace of change in fluorescence from baseline (F/F₀) of a single cultured dermal lymphatic endothelial cell (passage 3) loaded with 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate (DAF-FM) to detect NO production. DAF-FM is essentially nonfluorescent until it interacts with NO to produce a fluorescent benzotriazole. The time course shows the accumulation of this fluorescent benzotriazole and changes in slope associated with treatments and washouts. Apparent elevations in the slope were visible shortly after afobazole treatment or after treatment with positive controls for endothelium-derived NO production, acetylcholine (ACh) and bradykinin (BK). AU, arbitrary units. B: summarized data from 90 lymphatic endothelial cells that responded to afobazole. Data were analyzed by one-way ANOVA followed by Dunnett’s test, with all groups compared with baseline as control.