Figure 4H-J primary data

Figure 4H-J legend. 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”), 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 (Ser386), total IRF3, phosphorylated TBK1 (Ser172), total TBK1, and β-actin as a loading control. As a positive control for IRF3 and TBK1 activation/phosphorylation, A549 cells were treated with poly(I:C) for 6 and 48 hours before protein lysates were collected. Protein was isolated from treated BC3-IFN-βp-tdTomato reporter cells without sorting (“bulk”), or after sorting the lytic+casp-i+anti-IFN Abs sample based on tdTomato expression.Western blot for phosphorylated TBK1 and phosphorylated IRF3 for bulk lytic+casp-i+anti-IFN Abs 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.