JCI - Toll-like receptor 9–induced type I IFN protects mice from experimental colitis

Toll-like receptor 9–induced type I IFN protects mice from experimental colitis

Kyoko Katakura, Jongdae Lee, Daniel Rachmilewitz, Gloria Li, Lars Eckmann, Eyal Raz

J Clin Invest. 2005; 115(3):695–702 doi:10.1172/JCI22996

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Figure 4

TLR9-activated DNA-PK mediates activation of IRFs and type I IFN via MyD88. (A) BM-PDCs were treated with ISS-1 (10 μg/ml) and nuclear extracts were prepared. NF-κB was detected by EMSA; activation of IRFs, pSTAT1, and pERK, by Western blotting. Results represent 1 of 2 experiments. (B) BM-PDCs (10⁶/ml) from RAG1^–/– and DNA-PK^–/– mice (B6) were stimulated with ISS-1 (10 μg/ml). Cytokine levels in the supernatants were measured 24 hours later. (C) BM-MDCs (10⁶/ml) from RAG1^–/– and DNA-PK^–/– mice (B6) were stimulated with ISS-1 (10 μg/ml). Cytokine levels in the supernatants were measured 24 hours later. (D) BM-MDCs (10⁶/ml) from WT and MyD88^–/– mice were stimulated with 10 μg/ml ISS-1 (ISS) or 50 ng/ml LPS. Activation of DNA-PK was measured by an in vitro kinase assay using GST-p53 as a substrate, as described in Methods. Lysates from BM-MDC from RAG1^–/– and SCID mice were stimulated with ISS-1 (10 μg/ml) and then were subjected to immunoprecipitation with anti-ubiquitin, SDS-PAGE, and immunoblotting with anti–IRF-7. Ub-, ubiquitinated. (E) Proposed localization of DNA-PK in the TLR9 signaling pathway. Ub, ubiquitin.