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. Author manuscript; available in PMC: 2013 Mar 23.
Published in final edited form as: Immunity. 2008 Mar;28(3):381–390. doi: 10.1016/j.immuni.2008.02.002

Figure 1. A20 Inhibits MDP- and DAP-Dependent NFκB Transcriptional Activity.

Figure 1

(A–C) NFκB-dependent luciferase reporter assays of NOD2 or NOD1 transfected 293T cells. Indicated amounts of (A) NOD2, (B) NOD1, or (C) cAMP response element (CRE) plasmid trans-fected 293T cells were stimulated with MDP, Tri-DAP, or forskolin, respectively. Cells were also cotransfected with either 0 ng (black columns), 5 ng (gray columns), or 25 ng (white columns) of an A20 expression plasmid. Note the induction of NOD2-mediated NFκB transcriptional activity by MDP, the induction of NOD1-mediated NFκB by Tri-DAP, and the dose-dependent inhibition of these inductions by A20. By contrast, forskolin induced cAMP response element driven luciferase activity is not suppressed by A20. Data are reported in relative luciferase units (RLU) and are representative of three independent experiments.

(D and E) A20 is required for restricting MDP induced responses. ELISA analyses of secreted IL-1β and IL-6 from BMDCs. BMDCs from Tnfaip3+/+ (white columns) and Tnfaip3−/− (black columns) mice were stimulated with the indicated doses of MDP for 12, 24, or 36 hr (for IL-6 analyses) or 24 hr (for IL-1β analyses), after which supernatants were harvested and analyzed by ELISA for (D) IL-6 and (E) IL-1β secretion. Data are representative of three independent experiments, with standard deviation (bars) indicated.