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TBK1 subunits within the dimer complement each other to exert kinase activity. HA-tagged WT TBK1 or G217R TBK1 was coexpressed with Flag-tagged WT, D135N, S172A TBK1, or EGFP as a control in TBK1 knockout 293T cells (A). Cell lysates were prepared 24 h after transfection; immunoprecipitations were conducted with anti-Flag beads followed by kinase assays (KA) with recombinant IRF3 as the substrate at 30 °C for 40 min (B).
