Steady-state mRNA levels in PBMCs. Northern blot analysis of specific cytokine mRNA extracted from resting PBMCs at times indicated. Lane 1, unstimulated PBMCs; lane 2, ITF2357 only (50 nM); lane 3, LPS (10 ng/mL); lane 4, LPS in the presence of ITF2357 (50 nM). In addition, LPS-induced TNFα, IL-1 β, IFNγ, and IL-8 were measured in the supernatant by specific ELISA, and the percent inhibition of LPS-induced cytokines by ITF2357 is indicated under the respective bands and time points are indicated. ND = not determined. Shown are the results of a single donor. Similar results were obtained in a second donor.