Figure 10.

The production of nitric oxide (NO) by macrophages at 24 h after activation was determined by analyzing ${\text{NO}}_{{\text{2}}^{-}}$ levels in the cell culture medium. Mitomycin C-treated bone marrow-derived macrophages (BMDMs) were cultivated for 24 h before they were activated with the TLR2/1 agonist Pam3 (100 ng/mL) in combination with either (A) rIFN-γ at various concentrations ranging from 100 to 0.0015 ng/mL in 4-fold dilutions or (B) vdIFN-γ at concentratios ranging from 100 to 0.0015 ng/mL in 16-fold dilutions. Non-stimulated BMDMs and BMDMs that were stimulated with Pam3, rIFN-γ, or vdIFN-γ were used as the negative controls. Nitrite (${\text{NO}}_{{\text{2}}^{-}}$) levels were measured in the macrophage culture medium at 24 h post-stimulation using the Griess test. Experiments were performed in triplicates and were repeated two times with similar results. The bars represent the mean values of triplicates ± SEM from one representative experiment.