Pcmv5-human ELF4 has been described previously (15). The full length HIPK2 was subcloned to Pcmv7.1 and Pcmv3xHA vectors. The truncated HIPK2-NT fragment (1923 aa), HIPK2△NLS (795828 aa), and all point mutants of HIPK2 were generated by QuikChange site-directed mutagenesis kits (Stratagene) using the construct encoding wild-type protein as the template. Each mutation was confirmed by sequencing. The plasmids coding human STING, MAVS, and TBK1 have been described previously (59). For reporter assays, pISRE-luc (Stratagene) was used. Other promoters were cloned into pGL3-Basic Vector (Promega) with the following promoter regions: −1 to −155 for mouse IFN-β, −1 to −957 for mouse IFN-α1, −1 to 855 for mouse IFN-α2, −1 to −476 for mouse IFN-α4, and −1 to −1205 for mouse IFN-α8. All constructs were verified by sequencing. Point mutants of IFN-β reporter were generated by QuikChange site-directed mutagenesis kits (200518, Stratagene) and the construct encoding wild-type IFN-β–luc as the template. Each mutation was confirmed by sequencing.

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