Fig. 6

NIK requires its kinase function and phosphorylation at Thr 561 to potentiate STING activation in the DNA pathway. a IFN-β luciferase reporter assay in HEK 293T cells co-transfected with plasmids encoding STING and wild-type NIK, (NIK WT), kinase dead NIK (NIK KD), or a Thr 561 phospho-acceptor mutant NIK (NIK T561A). Data are means ± SEM of one experiment run in duplicates out of three independent experiments. b Semi-native PAGE immunoblot analysis of STING dimerization in HEK 293T cells co-transfected with HA-STING and Myc-WT, KD, or T561A NIK plasmids. Results are representative of two independent experiments. c Co-immunoprecipitation and immunoblot between Myc-WT, KD, or T561A NIK and Flag-WT, KD, or T561A NIK co-transfected in HEK 293T cells. Results are representative of three independent experiments. d Co-immunoprecipitation and immunoblot of HA-IRF3 with Myc-STING in the absence or presence of Flag-WT or T561A NIK co-transfected in HEK 293T cells. Results are representative of two independent experiments. e Co-immunoprecipitation and immunoblot of Myc-WT, KD, or T561A NIK with HA-STING co-transfected in HEK 293T cells. Results are representative of three independent experiments. p < 0.05 is considered significant