Modified Vaccinia Virus Ankara Triggers Type I IFN Production in Murine Conventional Dendritic Cells via a cGAS/STING-Mediated Cytosolic DNA-Sensing Pathway
Figure 6
Viral DNA replication is not required for MVA-induced IRF3 phosphorylation.
(A) GM-CSF-BMDCs (1×106) were pre-incubated with either PAA at 200 µg/ml or with mock control for 1 h, and then infected with MVA at a MOI of 3 for 1 h. Cells were washed and incubated with fresh medium with or without PAA. Cells were collected at 1, 4, 8, and 24 h post infection. Viral DNA was extracted and purified. Real-time PCR was performed with primers and TagMan probe specific for the vaccinia ribonucleotide reductase l4L gene. (B) Western blot analysis of BMDCs infected with MVA at a MOI of 10 in the presence of absence of PAA. Whole-cell lysates were prepared. Equal amount of proteins were subjected to SDS-PAGE and immunoblotting with anti-phosphoserine-396 of IRF3 or anti-IRF3. Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) was used as a loading control. “hpi”, hours post infection. “M”, mock infection control.