Figure 4F-G primary data

Fig 4. TBK1 and IRF3 are necessary but not sufficient to direct IFN-β expression, as they are activated even in cells with low IFN-β expression. BC3-IFN-βp-tdTomato reporter cells were treated with 20 ng/ml TPA for 48 hours to induce the lytic cycle (“lytic (+TPA)”). Where indicated, the cells were also treated with 10 μM of the pan-caspase inhibitor IDN-6556 (“casp-i”), 10 μM of the TBK1 inhibitor MRT76307 (“TBK1i”), 10 μM of the cGAS inhibitor G140 (“cGASi”), and/or a cocktail of antibodies against type I IFNs and their receptor (“anti-IFN Abs”) at 1:2000 dilution. Protein lysates were probed for phosphorylated IRF3 (Ser396), and β-actin as a loading control. Protein was isolated from treated BC3-IFN-βp-tdTomato reporter cells without sorting (“bulk”), or after sorting the lytic+casp-i sample based on tdTomato expression. Western blot for phosphorylated IRF3 for bulk lytic+casp-i treated samples and sorted tdTomato+ and tdTomato- samples were quantified by normalizing the density of phosphorylated protein to unphosphorylated protein bands. Phosphorylation levels are plotted relative to the bulk lytic+casp-i or lytic+casp-i+anti-IFN Abs treated samples. Images shown are representative of 3 replicates. One-Way ANOVA followed by Tukey’s post hoc multiple comparisons test.