Fig. 8
A peptide from MAVS potentiates the antiviral response in vitro and in vivo. a Characterization of the interaction domain between NLK and MAVS. HEK293T cells were transfected with the MAVS mutants and NLK, immunoprecipitated using the Flag antibody, and then subjected to immunoblot analysis. b Effect of NAMDAP on the interaction between NLK and MAVS. HEK293T cells were transfected with NLK and MAVS, MG132 was added to protect MAVS from degradation, and 20 µg/ml NAMDAP and SeV were added for 8 h before performing immunoprecipitation experiments. Immunoblot analysis was performed using the indicated antibody. c, d Effect of NAMDAP on the inhibitory effect of NLK on MAVS toward IFNB1 and RANTES gene transcription and the antiviral response. c HEK293T cells expressing NLK were incubated with increasing amounts of NAMDAP before performing real-time PCR experiments (n = 3). d HEK293T cells expressing NLK were incubated with VSV-GFP at an MOI of 0.0001 and increasing amounts of NAMDAP, followed by plaque assays (n = 2). e–g Effect of NAMDAP on the production of antiviral cytokines and the antiviral response in vitro. HEK293T cells were infected with VSV-GFP at an MOI of 0.01 followed by monitoring via FACS analysis (e). Scale bar is 500 µm. The supernatant was collected and added to Vero cells before performing the plaque assay (f) (n = 2). mRNA was isolated to determine the transcription level of IFNB1 (g) (n = 3). h, i Effect of NAMDAP on the antiviral response in vivo. Mice (12) were infected with VSV-GFP (2 × 108 pfu), after which 10 mg/kg NAMDAP or vehicle was injected three times every 4 h via the tail vein. Their sera were collected after 12 h and subjected to the plaque assay (g). Survival assay of mice (four for each) injected with or without NAMDAP. The survival of the mice was monitored for 2 weeks (h) (n = 4). NAMDAP: NLK-associated and MAVS-derived antiviral peptide. Data are representative of three independent experiments. Data are presented as the mean ± SEM. Statistical significance was analyzed by ANOVA or Student’s t-test (*p < 0.05, ***p < 0.001). Source data (a–d, f–h) are provided as a Source Data file
