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. Author manuscript; available in PMC: 2015 Apr 1.
Published in final edited form as: J Immunol. 2014 Sep 3;193(7):3726–3735. doi: 10.4049/jimmunol.1400210

FIGURE 2.

FIGURE 2

The N-terminal TLR9 proteolytic cleavage product negatively regulates its signaling. (A) The influence of the N-terminal cleavage product on TLR9 signaling. IL-6 production was measured in RAW macrophages expressing TM-GFP or Nterm-TM-GFP after stimulation for 12 h with LPS (80 ng/ml), CpG-DNA (1 μM), or titration with the indicated concentrations of CpG-DNA. IL-6 production was measured by ELISA. (B) TNF-α production by macrophages stably expressing TM-GFP or Nterm-TM-GFP exposed for 12 h to CpG-DNA (1 μM). (C) IL-6 production by wild-type immortalized macrophages stably expressing TM-GFP or Nterm-TM-GFP after stimulation with LPS (80 ng/ml) or CpG-DNA (1 μM) for 12 h. (D) NF-κB luciferase reporter assays of HEK293T cells expressing full-length TLR9 in the presence of the indicated expression plasmids were performed 12 h after stimulation with 1 μM CpG-DNA. *p < 0.05 (Student t test). Data are representative of at least three independent experiments (average ± SD).