Figure 5.

Analysis of expression of Egr2 and M1-associated markers (NOS2, TNF, IL-1β, Cox-2) and IL-10 in IFNγ-activated macrophages with knockdown of Egr2. Bone-marrow-derived macrophages (BMDMs) were transfected with siRNA cocktail for Egr2 [Egr2(si)] or control siRNA [C(si)] for 24 h as described in Materials and Methods, and after which the cells were used as unstimulated [C(si) and Egr2(si)] or activated with IFNγ for another 24 h-time period [C(si)+IFN and Egr2(si)+IFN] as in Figure 3. The cells were washed, mRNA was isolated and the expressions of Egr2 (A), NOS2 (B), TNF (C), IL-1β (D), Cox-2 (E) and IL-10 (F) were analyzed by real-time PCR as described in Materials and Methods similar as for Figure 2. In (A–F), mean ± S.E. of six separate culture plate wells is shown (^**p < 0.01; ^***p < 0.001; ^*****p < 0.00001).