Fig. 4

NOD2 mediates Cpn0423-induced cytokine production. Bone marrow-derived macrophages (BMDMs) were pretreated respectively with lipofectamine-complexed NOD2-siRNA or sterile water for 48 h before the administration of Cpn0423 (10 μg/ml), MDP (NOD2 ligand, 20 ng/ml), and Pam3Cys (P3C, TLR2 ligand, 10 ng/ml). mRNA and protein levels of NOD2 or TLR2 were assessed by quantitative real-time PCR and western blot respectively, and expression was normalized to that of GAPDH (a-c). Sixteen hours later, the culture media were collected and cytokine expression in the media was measured by MIP-2, IL-6 and TNF-α ELISA respectively (d-f). Each error bar represented mean ± SD. Two-tailed unpaired student t test was used in Fig. 4. a-e * P < 0.05, ** P < 0.01, *** P < 0.001. n = 3 in each group