Figure 4. TANK negatively regulates the activation of NF-κB and AP-1 as well as gene expression in response to TLR7 stimulation in macrophages.
(a) Peritoneal macrophages from wild-type (WT) and Tank−/− mice were stimulated with 10 nM R-848 for the indicated periods. Total RNA was extracted and subjected to Northern blot analyses for the expression of Il6, Tnf, Il12b, Ptgs2, Nfkbiz and Nos2. The same membranes were rehybridized with an Actb probe. Data of two independent experiments (lanes marked 1 and 2 represent distinct experiments) are shown. (b,c) Wild-type and Tank−/− macrophages were stimulated with R-848 (10 μM) for the indicated periods. Nuclear extracts were prepared, and the NF-κB (b) and AP-1 (c) DNA-binding activities were determined by EMSA using NF-κB- and AP-1-specific probes. The arrows indicate the induced NF-κB and AP-1 complexes. The results are representative of three independent experiments.