Fig. 1

The modulators of the alternative NF-κB pathway play a negative regulatory role in the DNA pathway. a, b IFN-β quantification via ELISA from supernatants collected 24 h postinfection or stimulation of WT and Traf3^−/− MEFs infected with Sendai virus (SeV) or herpes simplex virus 1 (HSV-1) (MOI 0.1) (a) or transfected with poly I:C (100 ng/mL) or B-DNA (500 ng/mL) (b). Data are means ± SEM of one experiment run in duplicates out of 2–3 independent experiments. c Immunoblot analysis of IRF3 activation in WT and Traf3^−/− MEF cells infected HSV-1 (MOI 0.1) at the indicated time points. Results are representative of two independent experiments. d, e Viral titers determined via plaque assay from supernatants of WT and Traf3^−/− MEFs infected with HSV-1 (MOI 0.1) (d) or vesicular stomatitis virus (VSV) (MOI 0.01) (e) analyzed 18, 21, and 24 h postinfection. Data are means ± SEM of one experiment run in duplicates out of 2–3 independent experiments. f Immunoblot analysis of HSV-1 encoded protein ICP4 in WT and Traf3^−/− MEF cells infected HSV-1 (MOI 0.1) at the indicated time points. Actin serves as a loading control. Results are representative of two independent experiments. g, h Gene expression levels of IFN-β determined via Q-PCR in WT and Traf2^−/− MEFs (g) or in WT and ciap1/2^−/− MEFs (h) stimulated with B-DNA (500 ng/mL) for 4 h. Data are means ± SEM of one experiment run in triplicates out of 2–3 independent experiments. p < 0.05 is considered significant