Fig. 1.

Effect of eNOS siRNA and L-NAME on cytokines: a HMVECs were treated with 72 h of siRNA scrambled (siControl) or eNOS (sieNOS) and then treated with Poly I:C (TLR3 agonist, 10 μg/mL) for 16 h. Supernatants were collected and the amount of IL-6, IP-10 and sVCAM produced among treatment groups was measured by ELISA (n = 4 per group). b HMVECs were exposed to L-NAME (100 μmol) or vehicle control (DMSO < 0.1%) 30 min prior to Poly I:C (10 μg/mL) treatment. Supernatants were then collected after 16 h of Poly I:C treatment and measured for IL-6 and IP-10 by ELISA (n = 4 replicates per group). c HMVECs were plated on transwells for 24 h. Baseline levels for normalization are shown at time 0. Cells were pretreated with L-NAME for 30 min and then Poly I:C afterwards at the doses listed in B. Change in trans-endothelial electrical resistance (TEER) from baseline (---) was determined as hourly measurements over 16 h from Poly I:C exposure (n = 4 replicates per group). * = p < 0.05 between compared groups, NS = non-significant