Fig. 2

NIK is a positive regulator of the DNA pathway. a Immunoblot analysis of TRAF3 and NIK in MEF cells (upper panel) or A549 cells (lower panel) transfected with B-DNA (2 μg/mL) for the indicated time points. Actin and HSP90 serve as loading controls. Results are representative of three independent experiments. b, c IFN-β mRNA levels determined via Q-PCR in WT and Nik^−/− MEFs (b) transfected with B-DNA (500 ng/mL) or in WT and Nik^−/− BMDMs (c) transfected with B-DNA (100 ng/mL) for the indicated time points. Data are means ± SEM of one experiment run in duplicates (b) or triplicates (c) out of 2–3 independent experiments. d ELISA for IFN-β in supernatants collected 24 h after infection from WT and Nik^−/− BMDMs infected with HSV-1 (MOI 5.0). Data are means ± SEM of one experiment run in duplicates out of two independent experiments. e, f Viral loads of HSV-1 encoding luciferase (e) or murine herpes virus 68 (MHV-68) encoding luciferase (f) in WT and Nik^−/− MEFs infected with virus at the indicated MOIs. Data are means ± SEM of one experiment run in triplicates out of two independent experiments. g Immunoblot analysis for IRF3 and TBK1 activation in WT and Nik^−/− BMDMs transfected with B-DNA (1 μg/mL) for the indicated time points. Results are representative of two independent experiments. p < 0.05 is considered significant