Fig. 3: WDR77 interacts with MAVS and inhibits its activity.
From: WDR77 inhibits prion-like aggregation of MAVS to limit antiviral innate immune response
a–c HEK293T cells were transfected with luciferase reporters and plasmids as indicated for 24 h. Cells were collected and IFNB1 (a), ISRE (b) and NFKB1 (c) promotor activation were detected by luciferase assay (For a, IFNB1: ****p < 0.0001; ****p < 0.0001; nsp = 0.9496 in sequence; For b, ISRE: ****p < 0.0001; ****p < 0.0001; nsp = 0.8583, nsp = 0.4715 in sequence; For c, NF-KB1: nsp = 0.2570 in sequence). d HEK293T cells were transfected with HA-WDR77, Flag-PRMT5, RIG-I, MAVS, TRAF2, TRAF3, TRAF5, TRAF6, TBK1, IRF3 or IKKβ-expressing plasmids as indicated for 36 h. Cells were collected and subjected to immunoprecipitation assay and immunoblotting. Asterisk indicated nonspecific bands. e Immunofluorescence assay of co-localization of HA-WDR77 (green) with Flag-MAVS (red) in HEK293T cells following SeV infection for 8 h. Nuclei were stained with DAPI (blue). Scale bars indicate 10 μm (Colocalization radio: ****p < 0.0001). f Diagram of MAVS truncations used in Co-IP experiments (top panel). Various MAVS truncations were co-expressed with Flag-WDR77 in HEK293T cells for 36 h before immunoprecipitation and immunoblotting (bottom panel) were performed. Asterisk indicates nonspecific bands. g Diagram of WDR77 truncations used in Co-IP experiments (top panel). Various WDR77 truncations were co-expressed with HA-MAVS in HEK293T cells for 36 h before immunoprecipitation and immunoblotting (bottom panel) were performed. h WT or WDR77-/- #1 HEK293T were transfected with various WDR77 truncations for 24 h, and then stimulated with or without SeV for 12 h. IFNB1 induction was measured by qPCR (IFNB1: all ****p < 0.0001; nsp = 0.9998). Data are representative of three independent experiments with similar results (d, e (left), f and g), or three independent experiments (a–c, e (right) and h) (mean ± SD of three biological replicates). P-values were determined by two-way ANOVA (Å Ãdák’s test) (a–c, h) or unpaired two-sided t-test (e). n.s. indicates no statistical significance. Source data are provided as a Source Data file.
