Fig. 5: PWWP3A translocates to the mitochondria and interacts with VISA.

a 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 anti-HA or anti-Flag. b Pull-down and immunoblot analysis of the direct interaction between purified PWWP3A proteins and indicated signaling molecules. 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 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. d Confocal microscopy analysis in HEK293 cells transfected with NYFP-PWWP3A and CYFP-VISA for 20 h. Scale bar, 20 μm. e The mitochondrial fraction (45%), cytoplasm fraction (20%), nucleus fraction (1%) or whole cell lysates (WCL) from the uninfected or SeV infected HEK293 cells were analyzed by immunoblot analysis with the indicated antibodies. f HEK293 cells were transfected with the indicated plasmids. Twenty hours later, co-IP was performed with control IgG or anti-Myc. The immunoprecipitates and lysates were analyzed by immunoblotting with anti-Myc or anti-Flag. g HEK293 cells were infected with SeV at the indicated time points, then co-immunoprecipitation was performed with control IgG or anti-PWWP3A. The immunoprecipitates and lysates were analyzed by immunoblotting with indicated antibodies. h HEK293 cells treated with KPT-185 (1 μM) for 4 h, followed by the SeV infection at the indicated time points. Then the cytoplasm fraction (20%) and nucleus fraction (1%) from the cells were analyzed by immunoblot analysis with the indicated antibodies. At least three independent experiments were performed with similar results. Source data are provided as a Source Data file.