pRK5 plasmids encoding hemagglutinin (HA)–tagged ubiquitin wild-type (wt), K48, and K63 were provided by T. Dawson (Addgene plasmids #17605, #17606, and #17608) (51). Flag-tagged IRF7 wt was purchased from GenScript, and the phosphomimetic Flag-IRF7 (S477D/S479D) was from John Hiscott Pasteur Laboratories (Istituto Pasteur Fondazione Cenci Bolognetti, Rome, Italy) (21). HA-Pin1–expressing plasmid was provided by H.-Y. Kao (Research Institute of University Hospitals of Cleveland, Cleveland, OH). Human full-length TRIM8 complementary DNA (cDNA) was isolated from human cDNA library using standard PCR techniques and was subsequently cloned into pcDNA3.1D/V5-His-TOPO plasmid (Invitrogen). The TRIM8 RING mutant C15S-C18S-C35S-C38S was generated by site-directed mutagenesis using the GeneArt Site-Directed Mutagenesis System (Thermo Fisher Scientific). Myc-GFP–tagged TRIM8 deletion mutants were provided by G. Meroni (Department of Life Sciences, University of Trieste, Trieste, Italy).

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