Huh7, HeLa and 293T cells were maintained in DMEM (Invitrogen) with 10% FCS and 0.1 mM non-essential amino acids. NIH-3T12 cells were grown in DMEM supplemented with 5% FCS, 100 U penicillin per ml, 100 μg streptomycin per ml and 2 mM L-glutamine.
STAT1−/− fibroblasts (an SV40 large T antigen immortalized human skin fibroblast line) were grown in RPMI (Invitrogen) with 10% FCS. The construction, characterization and generation of viral stocks for the following viruses have been previously described: CVB-GFP (derived from infectious clone pMKS1-GFP)
24 (link), PV-GFP (strain P1M, derived from infectious clone pPVM-2A144-GFP)
25 (link), EAV-GFP (derived from infectious clone pEAV211-GFP2aT)
26 (link), SINV-A-GFP and SINV-G-GFP (derived from infectious clones pS300-GFP and pG100-GFP)
27 (link), ONNV-GFP (derived from infectious clone pONNV.GFP)
28 (link), VEEV-GFP (derived from pTC83-GFP infectious clone)
4 (link), FLUAV-GFP (based on strain PR8)
29 (link), PIV3-GFP (based on strain JS)
30 (link), NDV-GFP (based on strain Hitchner B1)
31 (link), HMPV-GFP
32 (link) (based on isolate CAN97-83), RSV-GFP (based on strain A2)
32 (link), MV-GFP (MVvac2-GFP, based on vaccine strain, Edmonston lineage measles virus)
33 (link) and BUNV-GFP
34 (link) (based on rBUN-del7GFP). VV-GFP was propagated in BSC-40 cells. Viral stocks were prepared by three freeze–thaw cycles, followed by centrifugation at 1,000
g to remove cellular debris. VV Western Reserve was obtained from the ATCC, propagated in Vero cells and purified by ultracentrifugation through a 36% sucrose cushion. MHV68 clone WUMS was obtained from the ATCC and propagated in NIH-3T12 cells. The WNV strain was isolated and passaged as described previously
35 (link).
Schoggins J.W., MacDuff D.A., Imanaka N., Gainey M.D., Shrestha B., Eitson J.L., Mar K.B., Richardson R.B., Ratushny A.V., Litvak V., Dabelic R., Manicassamy B., Aitchison J.D., Aderem A., Elliott R.M., García-Sastre A., Racaniello V., Snijder E.J., Yokoyama W.M., Diamond M.S., Virgin H.W, & Rice C.M. (2013). Pan-viral specificity of IFN-induced genes reveals new roles for cGAS in innate immunity. Nature, 505(7485), 691-695.