Cell culture and gene knockout protocols

HEK293T cells were maintained in DMEM (Gibco) supplemented with 10% fetal calf serum (FCS, Labtech) and 100 U/ml penicillin and 100 μg/ml streptomycin (Pen/Strep; Gibco). THP-1 cells were maintained in RPMI (Gibco) supplemented with 10% FCS and Pen/Strep. THP-1-IFIT-1 luciferase reporter cells express Gaussia luciferase under the control of the endogenous IFIT1 promoter have been described (Mankan et al., 2014 (link)). THP-1 CRISPR control, cGAS-/- and MAVS -/- knock out cells have been described (Mankan et al., 2014 (link)). Nup358 depleted HeLa cells have been described (Schaller et al., 2011 (link)). Lipopolysaccharide, poly I:C and TNFα were obtained from PeproTech. Sendai virus was obtained from Charles River Laboratories. Herring-testis DNA was obtained from Sigma. cGAMP was obtained from Invivogen. NF-ĸB Lucia THP-1 reporter cells were obtained from Invivogen. All cell lines were tested negative for mycoplasma.