Fig. 5: SAMD9 restricts the infection of vaccinia virus, rotavirus, and reovirus.

A EMSA of SAMD9 with rotavirus (RV) segmented dsRNA genome. B Nuclear, membrane, and cytoplasmic fractions from A549 cells prepared by homogenization and differential centrifugation. Pellet (P8000) and supernatant (S8000) from centrifugation of 8000×g immunoblotted with indicated antibodies. C A549 cells infected with mock or RV (MOI = 300) for 8 h, then fixed and stained for SAMD9 (green), dsRNA (red), nuclei (blue) and colocalization (yellow) by confocal microscopy. D A549 cells infected with mock or RV (MOI = 3) for 8 h and cell lysates harvested for native-PAGE or SDS-PAGE electrophoresis with indicated antibodies. E, F WT, SAMD9 KO, SAMD9 KO complemented with SAMD9-mGFP, RIG-I KO, RIG-I SAMD9 double KO, MDA5 KO, MDA5 SAMD9 double KO, RIG-I MDA5 double KO, RIG-I MDA5 SAMD9 triple KO, MAVS KO, and MAVS SAMD9 double KO HT-29 cells infected by RV (MOI = 0.01) for 24 h, the IFNL3 mRNA level (E) or CCL5 mRNA level (F) measured by qRT-PCR. G, H WT and SAMD9 KO complemented with Ev or SAMD9-mGFP (Rescue) HT-29 cells infected with RV (MOI = 0.01) for 24 h, the levels of secreted IFN-λ3 (G) and CCL5 (H) quantified by ELISA. I WT and SAMD9 KO Hela cells infected by indicated viruses (MOI = 0.1) for 24 h and the mRNA levels of viral genes measured by qRT-PCR. RRV rhesus rotavirus, UK bovine rotavirus, WI61 human rotavirus, Reo mammalian orthoreovirus, IAV influenza A virus, VSV vesicular stomatitis virus, CHIKV chikungunya virus, ZIKV Zika virus, HAstV human astrovirus, AdV adenovirus. For all figures, experiments were repeated at least three times. Individual points in (E–I) represent independent experiments. Data are represented as mean ± SEM. Statistical significance calculated by two-way ANOVA with Tukey’s multiple-comparisons tests (E–I): n.s., not significant, *P < 0.05, **P < 0.01, ***P < 0.001. Source data are provided as a Source Data file. Detailed P-values are provided in the Source Data file.