Fig. 7: PWWP3A competes with TRAF6 to bind to VISA.

a Protein–protein interaction (PPI) map for PWWP3A and immune response related interactors. b HEK293 cells were transfected with the indicated plasmids for 20 h before the cells were lysed and centrifuged, then the supernatants were incubated with E. coli-derived His-PWWP3A at 37 °C for 2 h before pull-down assays were performed. c HEK293 cells were infected with sendai virus (SeV) at the indicated time points, then co-immunoprecipitation was performed with control IgG or anti-TRAF6. The immunoprecipitates and lysates were analyzed by immunoblotting with indicated antibodies. d HEK293 cells were transfected with the indicated plasmids. Twenty hours later, co-IP was performed with control IgG or anti-HA. The immunoprecipitates and lysates were analyzed by immunoblotting with the indicated antibody. e The Flag-VISA expressed cell lysate was incubated with E. coli-derived GST-TRAF6 at 37 °C for 2 h before co-IP analysis. f The control and PWWP3A-knockout (KO) HEK293 cells were infected with SeV at the indicated time points, then co-immunoprecipitation was performed with control IgG or anti-VISA. The immunoprecipitates and lysates were analyzed by immunoblotting with indicated antibodies. g, h HEK293 cells were transfected with the indicated plasmids. Twenty hours later, co-IP was performed with control IgG or anti-Flag. The immunoprecipitates and lysates were analyzed by immunoblotting with the indicated antibody. i The pull-down assay was performed as described in (b) except HA-VISA and its mutant (VISA-d6) were used here. VISA-d6, the mutant of VISA in which both the E155 and E457 of VISA were mutated to aspartic acid (D). At least three independent experiments were performed with similar results. Source data are provided as a Source Data file.