Dengue virus NS1 cytokine-independent vascular leak is dependent on endothelial glycocalyx components
Hair was removed from the dorsal dermis of wild-type B6 mice, and mice were allowed to recover for 3 days. On the day of the assay, mice received an intraperitoneal dose of inhibitor cocktail (Zanamivir, Cathepsin L Inhibitor, and OGT 2115; 1 mg/ml of each inhibitor) 6 hours pre-assay and then immediately preceding the start of the assay (n = 4; closed symbols). Control mice received injections of DMSO, PBS, and water as a vehicle control (n = 3; open symbols). Retro-orbital injections of Alexa Fluor 680-conjugated dextran were then administered, followed by intradermal injections of PBS (black circles), 200 ng VEGF (purple squares), 15 μg DENV2 NS1 (blue triangles), and 7.5 μg DENV2 NS1 (green triangles). The dermis from each mouse was collected and processed two hours post-injection. Data represent the fold change of mean fluorescent intensity from VEGF and DENV2 NS1 injections to PBS injections. Data represent mean +/- SEM and were collected from 2 independent experiments. Unpaired, parametric, two-tailed t-tests were used to determine significance between inhibitor-treated and mock-treated groups. ns = not significant, *P < 0.05, **P < 0.01.