Fig. 2: SAMD9 senses cytosolic dsDNA and dsRNA.

A Validation of WT, SAMD9 KO, SAMD9-mGFP add-back SAMD9 KO, STING KO, STING SAMD9 double KO HT-29 cells by western blot with indicated antibodies. B WT, SAMD9 KO, SAMD9 KO complemented with SAMD9-mGFP, STING KO, and STING SAMD9 double KO HT-29 cells transfected with poly(dA:dT) (1 μg/ml) for 24 h and the IFNL3 mRNA level measured by qRT-PCR. C Validation of WT, SAMD9 KO, SAMD9-mGFP add-back SAMD9 KO, 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, MAVS SAMD9 double KO HT-29 cells by western blot with indicated antibodies. D 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 transfected with LMW poly(I:C) (1 μg/ml) for 24 h and the IFNL3 mRNA level measured by qRT-PCR. E, F Validation of WT and two individual single clonal SAMD9 KO human intestinal organoids by western blot (E) and immunofluorescence staining F. G WT and two individual SAMD9 KO clones of human ileal organoids transfected with poly(dA:dT) or LMW poly(I:C) (1 μg/ml) for 24 h and the IFNL3 mRNA level measured by qRT-PCR. Human ileal organoids fixed and stained for SAMD9 (green) and nuclei (blue) by microscopy. H Validation of WT and SAMD9 KO primary human hepatocytes by western blot. I IFNL3 mRNA levels in poly(dA:dT) or LMW poly(I:C) (1 μg/ml) transfected WT and SAMD9 KO primary human hepatocytes measured by qRT-PCR. For all figures, experiments were repeated at least three times. Individual points in (B, D, G, and I) represent independent experiments. Data are represented as mean ± SEM. Statistical significance calculated by two-way ANOVA with Tukey’s multiple-comparisons tests (B, D, G, and 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.