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. Author manuscript; available in PMC: 2013 Oct 1.
Published in final edited form as: Nat Immunol. 2013 Mar 3;14(4):327–336. doi: 10.1038/ni.2548

Figure 1. TRIM21 senses intracellular Ab-bound virus.

Figure 1

(a) DNA binding ELISA showing NF-κB subunits p65 and p50 binding to consensus oligonucleotides 4 h post challenge of wild-type (WT) MEFs or Trim21-deficient MEFs transduced with empty vector (K21 EV) or expressing human Trim21 (K21 T21) with PBS, anti-adenovirus monoclonal antibody 9C12 (Ab), adenovirus (AdV) or adenovirus-antibody complex (AdV + Ab). (b) Induction of NF-κB luciferase reporter activity in wild-type or Trim21-deficient (K21) MEFs 7 h after challenge with a serial dilution of goat anti-adenovirus, AdV or adenovirus-antibody complex over PBS-treated controls. (c) Induction of NF-κB luciferase reporter activity in wild-type, Trim21-deficient and Trim21-deficient cells expressing human TRIM21. (d) Induction of NF-κB luciferase reporter activity in HeLa cells or HeLa cells depleted of TRIM21 by siRNA. Ab is pooled human serum IgG. (e) Induction of NF-κB luciferase reporter activity after challenge of Trim21-deficient MEFs transduced with empty vector or human TRIM21 with AdV, human serum IgM or AdV + IgM complex. (f) Hot-spot interactions between IgG Fc (cyan) and TRIM21 PRYSPRY domain (yellow) required for complex formation (based on PDB structure 2IWG). (g) Induction of NF-κB luciferase reporter activity in wild-type MEFs challenged with AdV incubated with recombinant 9C12 (r9C12) or point mutant N434D (r9C12 ND). (h) Induction of NF-κB luciferase reporter activity in AdV non-permissive EL4 cells and AdV permissive EL4 CAR 7 h after challenge. For panels a-e, g, h error bars represent SEM from three replicates.