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. 2011 Sep 26;208(10):2099–2112. doi: 10.1084/jem.20102667

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© 2011 Yang et al.

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Figure 2. — Silencing of CHIP impairs TLR signaling pathways. (A) RAW264.7 cells were transfected with plasmids encoding siRNAs, and selected with 600 µg/ml neomycin. The efficiency of silencing was evaluated by Western blot. CTRL, scrambled control RNAi; RNAi, CHIP-specific RNAi. (B and C) Reporter assays in RAW264.7 cells. Cells in A were transiently transfected with NF-κB or IRF3 reporters (B) or IL-6, CCL5, or IFN-β promoter reporters for 48 h, and then treated with 100 ng/ml LPS, 10 µg/ml poly (I:C), or 1 µM CpG ODN for 4 h. Results were expressed as fold induction relative to the luciferase activity in unstimulated cells transfected with control vector. Data are shown as mean ± SD of triplicate samples. ns, not significant; *, P < 0.05; **, P < 0.01. The data shown correspond to a representative experiment out of three performed.