Figure 4.

Analysis of expression of Egr2 and M2-associated markers (Arg1, Fizz1, Ym1, PPARγ) and IL-10 in IL-4-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 IL-4 for another 24 h-time period [C(si)+IL4 and Egr2(si)+IL4] as in Figure 3. The cells were washed, mRNA was isolated and the expressions of Egr2 (A), Arg1 (B), Fizz1 (C), Ym1 (D), PPARγ (E), and IL-10 (F) were analyzed by real-time PCR as described in Materials and Methods. In (A–F), mean ± S.E. of 4–9 separate culture plate wells is shown (^*p < 0.05; ^**p < 0.01; ^***p < 0.001; ^****p < 0.0001; ^*****p < 0.00001).