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. 2016 Jul 5;7:1003. doi: 10.3389/fmicb.2016.01003

FIGURE 5.

FIGURE 5

Involvement of TLR4 in the classical activation of macrophages by paracoccin. Macrophages (1 × 106 cells/mL) from WT, TLR2-/-, and TLR4-/- were stimulated for 6 h with p-rPCN (5 μg/mL). As a positive control for the classical activation, the mixture of IFN-γ (1 ng/mL) and IL-12p40 (50 ng/mL) was used. For alternative activation, a mixture of IL-10 plus IL-4 (50 ng/mL both) was used. The medium alone was used as the negative control. After extraction using TRizol Reagent®, the RNA was converted into cDNA and the expression of iNOS (A), Arginase-1 (B), Ym-1 (C), and FIZZ1 (D) were analyzed by real-time PCR. The relative expressions were determined as described in Section “Materials and Methods” and compared between WT, TLR2-/- and TLR4-/- macrophages. The results are expressed in mean ± SEM and were compared through one-way analysis of variance, followed by Bonferroni’s test. ∗∗∗p < 0.001, ∗∗p < 0.01, ∗p < 0.05, and non-significant differences (n.s).