Human a549 lung carcinoma cell line

Murine embryonic fibroblasts (MEFs) were derived from wild type (WT) C57BL/6, TLR3/IPS-1^−/− and TLR3/IPS-1/MDA5^−/− mice and were maintained in α-MEM supplemented with 10% FBS, 100 U ml⁻¹ penicillin, 100 μg ml⁻¹ streptomycin and 2mM L-glutamine. The A549 human lung carcinoma cell line (American Type Culture Collection) was maintained in α-MEM supplemented with 10% FBS. Vero cells (ATCC) were maintained in DMEM supplemented with 5% FBS. All cells were incubated at 37°C in a humidified 5% CO₂ incubator.
Polyinosinic/polycytidylic acid (poly IC) was purchased from GE Healthcare (Buckinghamshire, UK). Doxycycline hyclate and oligomers were purchased from Sigma-Aldrich, Canada. Puromycin was purchased from Gibco (life technologies, USA). The PI3K inhibitor wortmannin (novex, life technologies) was reconstituted in DMSO. Human recombinant platelet-derived growth factor (PDGF-BB; Gibco) was reconstituted in PBS containing 0.1% BSA at a concentration of 50 ng/μl. Real-time PCR Taqman probes for human ISG56, ISG15 and GAPDH were purchased from Applied Biosystems (Streetsville, Canada).

Mouse Lewis Lung Carcinoma cells (LLC, CRL-1642), H1299 human lung carcinoma cell line (CRL-5803), H358 bronchioalveolar carcinoma cell line (CRL-5807) and A549 human lung carcinoma cell line (CCL-185) were purchased from the American Type Culture Collection. The cells were not specifically characterized for this study. DMEM, RPMI-1640 and Ham's F12 Nutrient Mixture were from Mediatech, Inc., (Manassas Herndon, VA). Fetal bovine serum (FBS) was purchased from Equitech-bio, Inc. (Kerrville, TX). Penicillin-streptomycin and trypsin-EDTA were from Invitrogen (Carlsbad, CA). Other chemicals were analytical grade.