Temporal pattern of ET-induced IL-6 secretion, NO production, and IL-6 and iNOS mRNA expression. Subconfluent SCp2 cells were treated with ET (0 or 10 μg/ml) in 1% FBS DM as described in Methods. (A) IL-6 secretion assayed by ELISA and (B) NO production assayed by Griess reaction. RNA harvested at each time point were analyzed by RT-qPCR for ET-induced (C) IL-6 mRNA and (D) iNOS mRNA and quantified relative to GAPDH mRNA for each. Solid-line represents the results for control non ET-treated cells; while dashed-line represents the results for ET-treated cells. Each experiment was performed at least 3 times. Samples were assayed separately in duplicate analyses by either ELISA or Griess reaction assay. For RT-qPCR, each sample was analyzed in triplicate RT-qPCR reactions. Data represents the average of samples from 3 experiments ± SEM with (*) denoting significant differences from non-ET control within each time point (p < 0.05).