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. Author manuscript; available in PMC: 2014 Jul 18.
Published in final edited form as: J Immunol. 2010 Nov 1;185(11):6599–6607. doi: 10.4049/jimmunol.1002041

FIGURE 5.

FIGURE 5

Mapping of critical LPS, IFN-α, and IL-27 response elements in IL-10 promoter by substitution mutant analysis. A, 6-nt mutants were constructed spanning across entirety of the regions −1541/−1232 and −595/−455 within the context of the −1954/+64 IL-10 promoter reporter construct. Constructs were transfected in RAW 264.7 cells and assessed for luciferase activity as described in B. Sequences of representative wild-type or substitution mutants are indicated. B, Empty pGL4.20, −1954/+64 IL10 promoter, and mutant IL-10 promoter constructs (−1954/+64 backbone) were transfected into RAW264.7 cells and left unstimulated or stimulated with LPS (1 µg/ml) (B), rIFN-α (1000 U/ml) (C), and rIL-27 (80 ng/ml) (D). Luciferase activity is displayed as percent of full-length wild-type (−1954/+64) IL-10 promoter activity normalized to renilla activity representing three independent experiments. Student t test was performed. *p < 0.05.