FIGURE 1.

Ac₂PIM differentially regulates NOD2-induced immune response. A, peritoneal macrophages were treated with Ac₂PIM followed by MDP for the indicated time points. Lysates were analyzed for pRIP2 and pTAK1 by immunoblotting. B-D, immunoblot analysis of COX-2, SOCS-3, and MMP-9 was performed on lysates obtained from macrophages after the following treatments: treatment with pharmacological inhibitor PP2 prior to 12 h MDP addition (B), treatment with indicated concentrations of Ac₂PIM followed by 12 h treatment of MDP (C), and pretreatment of macrophages from C57BL/6J WT and tlr2-KO mice with Ac₂PIM followed by MDP for 12 h (D). E–H, macrophages were pretreated with indicated concentration of Ac₂PIM followed by MDP for 12 h. Cell-free supernatants were analyzed for NO by Greiss reagent (E), TNF-α (F), VEGF-A (G), and IL-12 (H) by ELISA. All data represent the mean ± S.E. from three independent experiments, ***, p < 0.001 (one-way ANOVA) and all blots are representative of three independent experiments. The cells were treated with 2 μg/ml Ac₂PIM unless mentioned otherwise for 2 h followed by 200 ng/ml MDP. Med, medium; WT, wild-type; KO, knock-out; DMSO, dimethyl sulfoxide.