Figure 8.

Characterization of TNF-α production induced by Compounds 1 and 2 in murine macrophages. Panel A: J774.A1 macrophages (2×10⁵ cells/well) were pretreated for 30 min with control buffer (0) or PTX (0.4 and 1 µg/ml), followed by the addition of Compound 1 (50 µM), 2 (50 µM), or LPS (50 ng/ml) for 12 hr, and TNF-α was measured in the cell supernatants by ELISA. Statistically significant differences (*P<0.001) between responses induced in PTX-pretreated versus DMSO-pretreated controls are indicated. Panel B: Macrophages (2×10⁵ cells/well) were cultured in the presence of control DMSO, fMLF (50 µM), WKYMVm (50 µM), or Compound 1 (25 µM) for 24 hr, and TNF-α was measured in the cell supernatants by ELISA. Panel C: Macrophages (2×10⁵ cells/well) were pretreated for 5 min with control DMSO, Boc-2 (40 µM), or WRW4 (10 µM), followed by the addition of Compound 1 (50 µM), 2 (50 µM), or LPS (50 ng/ml) for 24 hr, and TNF-α was measured in the cell supernatants by ELISA. Statistically significant differences (*P<0.001) between responses induced by LPS or Compounds 1 and 2 in Boc-2- or WRW4-pretreated versus vehicle-pretreated controls are indicated. The data in all panels are presented as the mean±SD of triplicate samples from one experiment, which is representative of three independent experiments.