FIGURE 6.

Impact of TRAF6 arginine methylation on the ligand-induced TLR response. A, THP-1-Lucia NF-κB reporter monocytes were transfected with negative control siRNA or JMJD6-specific siRNA and treated with ligands for different TLR molecules as indicated. Results of luciferase reporter assays are presented as the mean ± S.D. *, p < 0.05; **, p < 0.01 compared with negative control (NC) siRNA. n ≥ 3. B, luciferase reporter assays in Huh7.5 cells (left panel) or THP-1-Lucia NF-κB reporter monocytes (right panel) transfected with negative control siRNA or PRMT1-specific siRNA. Cells were treated with TLR ligands as indicated. Data are presented as the mean ± S.D. †, p < 0.05; ††, p < 0.01 compared with negative control siRNA. n ≥ 3. C, -fold increase in mRNA levels induced by FSL-1 exposure was determined in Huh7.5 cells treated with or without PRMT1 inhibitor AMI-1 for 16 h where indicated. *, p < 0.05; **, p < 0.01 compared with control; †, p < 0.05 compared with FSL-1 only. ICAM, intercellular adhesion molecule 1. D, FSL-1-induced -fold mRNA changes are shown in primary peritoneal macrophages. Cells were transfected with negative control (NC) siRNA and PRMT1-specific or JMJD6-specific siRNA where indicated. *, p < 0.05, compared with control; †, p < 0.05 compared with NC siRNA, n = 3. E, NF-κB luciferase reporter assays in Huh7.5 cells in the presence or absence of TLR6/2 ligand (1 ng/ml of FSL-1) that were transfected with negative control siRNA, PRMT1-specific siRNA, or both PRMT1- and JMJD6-specific siRNA. Data are presented as the mean ± S.D. *, p < 0.05; **, p < 0.01 compared with NC siRNA; ††, p < 0.01 compared with P1 siRNA. n = 3.