Fig 2: Propranolol treatment inhibited the IFN-I-mediated induction of antiviral genes and counteracted IFN-mediated inhibition of viral propagation in HGC-27 human gastric cancer cells. A and B Dose effects of propranolol were evaluated on the IFN-induced inhibition of T1012G production in HGC-27 cells. Cells were treated with 300 ng/well of human recombinant IFN-α/β and different concentrations (0, 10, 20, or 40 μmol/l) of propranolol for 48 h and infected withT1012G (0.01 MOI). Two days after infection, the cells and medium were harvested, and yields of virus were determined were determined on Vero cells. C and E HGC-27 cells were incubated with 40 μmol/l propranolol (for 48 h) and/or 300 ng/well human recombinant IFN-α/β (for 6 h) and then harvested for the assessment of protein expression of STAT3, p-STAT3, PKR, and p-PKR by western blot analysis. D and F Quantification of C, E. Results are presented as mean ± SEM, significant differences were evaluated using one-way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001(Dunnett’s test for multiple comparisons)

Fig 3: V117L and T314A UNC93B1 variants spontaneously induce IFN and NF-κB signaling pathways.a, RT–qPCR analysis of IFNβ, IFIT3 and ISG-15 expression in the indicated THP-1 cell lines (n = 3 biological replicates). There were three independent experiments. b, Levels of phosphorylated NF-κB, JNK, p38 and IRF5, as measured by immunoblotting, in lysates of the indicated THP-1 cells. Data are representative of three independent experiments. c, Production of IL-6, IFNα and IFNβ in the indicated THP-1 cell lines (n = 3, three independent experiments measured by ELISA). The indicated P values in a and c were determined using unpaired, two-tailed Student’s t-test; the data are presented as the mean with s.d. d,e, RNA-seq was performed for UNC93B1V117L (d) and UNC93B1T314A (e) THP-1 cells compared with WT. DEGs are presented as a volcano plot and mRNA was extracted from THP-1 of UNC93B1WT, UNC93B1V117L and UNC93B1T314A (n = 3 biological replicates). f, RT–qPCR analysis of IL-6, IL-8 and TNF expression in the indicated immortalized B cell lines (baseline) (n = 3 biological replicates). g, Production of IL-6, IL-8 and IL-12p70 in the indicated immortalized B cell lines (baseline) (n = 3 biological replicates, measured by CBA). The indicated P values in f and e were determined by two-way ANOVA, multiple comparison, Padj value; data are presented as mean with s.d.Source data

Fig 4: REXO2 (T132A) spontaneously induces innate activation of IFN-I.A Expression of IFNA4 (Ctrl vs WT P = 0.0019, Ctrl vs T132A P < 0.0001), IFNAR2 (Ctrl vs WT P = 0.0009, Ctrl vs T132A P < 0.0001), IFNAR1 (Ctrl vs WT P = 0.1308, Ctrl vs T132A P < 0.0001), IFNB1 (Ctrl vs WT P < 0.0001, Ctrl vs T132A P < 0.0001), ISG15 (Ctrl vs WT P = 0.0805, Ctrl vs T132A P < 0.0001), ISG20 (Ctrl vs WT P = 0.0036, Ctrl vs T132A P < 0.0001), ISG20L2 (Ctrl vs WT P = 0.0528, Ctrl vs T132A P < 0.0001), CCL5 (Ctrl vs WT P = 0.1788, Ctrl vs T132A P < 0.0001), TNF (Ctrl vs WT P = 0.1027, Ctrl vs T132A P < 0.0001), ISG54 (Ctrl vs WT P = 0.0003, Ctrl vs T132A P < 0.0001), ISG56 (Ctrl vs WT P = 0.0007, Ctrl vs T132A P < 0.0001) and IL6 (Ctrl vs WT P = 0.0286, Ctrl vs T132A P = 0.0002) was measured by qPCR analysis of inflammatory signaling pathways related to type I IFN in vector alone (Ctrl), REXO2-FLAG (WT) and REXO2-FLAG (T132A) THP-1 stable cell lines. Data are mean ± SD, pooled from 6 independent experiments. The p values were calculated using two-way ANOVA to Ctrl. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, NS not significant. Adjusted p value. B Heat map showing IFN regulated genes in REXO2 (T132A) compared with REXO2 (WT) THP-1 stable cell lines (n = 3). C Phosphorylated IRF3 (pIRF3) and total IRF3, phosphorylated TBK1 (pTBK1) and total TBK1 expression in vector alone (Ctrl), REXO2 (WT) and REXO2 (T132A) THP-1 stable cell lines by western blot (representative blots are shown of three independent repeats). D Quantification of IFNβ (Ctrl vs WT P < 0.0001, Ctrl vs T132A P < 0.0001) by ELISA in vector alone (Ctrl), REXO2 (WT) and REXO2 (T132A) THP-1 stable cell lines. Data are mean ± SD, pooled from 9 independent experiments. The p values were calculated using two-way ANOVA to Ctrl. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, NS not significant. Adjusted p value. Source data are provided as a Source data file.

Fig 5: Inflammatory signaling from REXO2 (T132A) is dependent on MDA5/MAVS.A Expression of IFNB1 (NC Ctrl vs NC WT P = 0.0361, NC Ctrl vs NC T132A P = 0.0003, RIG-I sgRNA Ctrl vs RIG-I sgRNA WT P = 0.0021, RIG-I sgRNA Ctrl vs RIG-I sgRNA T132A P = 0.0003, PKR sgRNA Ctrl vs PKR sgRNA WT P = 0.0028, PKR sgRNA Ctrl vs PKR sgRNA T132A P < 0.0001), TNF (NC Ctrl vs NC T132A P < 0.0001, RIG-I sgRNA Ctrl vs RIG-I sgRNA T132A P < 0.0001, PKR sgRNA Ctrl vs PKR sgRNA T132A P < 0.0001), CCL5 (NC Ctrl vs NC T132A P = 0.0025, RIG-I sgRNA Ctrl vs RIG-I sgRNA T132A P < 0.0001, PKR sgRNA Ctrl vs PKR sgRNA T132A P = 0.0010), ISG15 (NC Ctrl vs NC WT P = 0.1752, NC Ctrl vs NC T132A P = 0.0045, RIG-I sgRNA Ctrl vs RIG-I sgRNA T132A P < 0.0001, PKR sgRNA Ctrl vs PKR sgRNA T132A P = 0.0032), ISG20 (NC Ctrl vs NC WT P = 0.0053, NC Ctrl vs NC T132A P = 0.0005, RIG-I sgRNA Ctrl vs RIG-I sgRNA WT P < 0.0001, RIG-I sgRNA Ctrl vs RIG-I sgRNA T132A P < 0.0001, PKR sgRNA Ctrl vs PKR sgRNA WT P = 0.0003, PKR sgRNA Ctrl vs PKR sgRNA T132A P < 0.0001), ISG20L2 (NC Ctrl vs NC WT P = 0.0039, NC Ctrl vs NC T132A P = 0.0004, RIG-I sgRNA Ctrl vs RIG-I sgRNA WT P < 0.0001, RIG-I sgRNA Ctrl vs RIG-I sgRNA T132A P < 0.0001, PKR sgRNA Ctrl vs PKR sgRNA WT P = 0.0154, PKR sgRNA Ctrl vs PKR sgRNA T132A P = 0.0276), ISG54 (NC Ctrl vs NC T132A P = 0.0037, RIG-I sgRNA Ctrl vs RIG-I sgRNA T132A P < 0.0001, PKR sgRNA Ctrl and PKR sgRNA T132A P = 0.0014), ISG56 (NC Ctrl vs NC WT P = 0.0502, NC Ctrl vs NC T132A P = 0.0001, RIG-I sgRNA Ctrl vs RIG-I sgRNA WT P < 0.0001, RIG-I sgRNA Ctrl vs RIG-I sgRNA T132A P < 0.0001, PKR sgRNA Ctrl vs PKR sgRNA WT P = 0.0006, PKR sgRNA Ctrl vs PKR sgRNA T132A P < 0.0001) and IL6 (NC Ctrl vs NC T132A P < 0.0001, RIG-I sgRNA Ctrl vs RIG-I sgRNA T132A P = 0.0004, PKR sgRNA Ctrl vs PKR sgRNA T132A P = 0.0002) was measured by qPCR analysis of inflammatory signaling pathways related to type I IFN in vector alone (Ctrl), REXO2 (WT) and REXO2 (T132A) THP-1 stable cell lines which were also transduced with a sgRNA vector (NC), MDA5 sgRNA, RIG-I sgRNA or PKR sgRNA. Data are mean ± SD, pooled from 3 independent experiments. The p values were calculated using one-way ANOVA to Ctrl. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, NS not significant. Adjusted p value. B Western blot of phosphorylated IRF3 (pIRF3), total IRF3, phosphorylated TBK1 (pTBK1) and total TBK1 in vector alone (Ctrl), REXO2-FLAG (WT), and REXO2-FLAG (T132A) THP-1 stable cell lines which were also transduced with a sgRNA vector (WT) or MDA5 sgRNA (representative blots are shown of three independent repeats). Source data are provided as a Source data file.