Innate Immune Response to Virus Infection

Non- and PUUV-Suo infected Mygla.REC.B cells were exogenously stimulated with TLR-3 ligand polyI:C (10 µg/mL, Sigma-Aldrich) or Sendai virus (multiplicity of infection 1; received from Prof. Ilkka Julkunen; National Institute of Health and Welfare, Helsinki, Finland). Cells were collected 1-day post-treatment, and RNA was extracted from cell cultures with Trisure (Bioline, London, UK) following the manufacturers’ instructions. Extracted RNA was then reverse transcribed to complementary DNA (cDNA) using random hexamers and RevertAid reverse transciptase (Thermo Scientific). Relative quantitative PCR was performed with Maxima SYBR Green master mix (Thermo Scientific) using AriaMx instrumentation (Agilent, Palo Alto, CA, USA). Published primer sequences were used to measure the mRNAs levels of the interferon-inducible gene were Mx2, a marker of innate immune activation towards virus infection [22 (link)], and actin, which is commonly used for normalization of mRNA levels between samples [23 (link)]. Fold changes of individual mRNA expression levels relative to mock-infected controls were performed by the comparative CT method [24 (link)].