Journal: Pathogens
Article Title: Interactome Profiling of N-Terminus-Truncated NS1 Protein of Influenza A Virus Reveals Role of 14-3-3γ in Virus Replication
doi: 10.3390/pathogens11070733
Figure Lengend Snippet: Examination of IFN-β mRNA expression in 14-3-3γ knockdown cells infected with influenza A virus. A549 cells were transfected with 14-3-3γ siRNA for 24 h and then infected with influenza A PR8 wild-type virus at an MOI of 2. At 3, 6, 9, and 12 h post-infection (PI), the total RNA of the infected cells was extracted to determine IFN-β mRNA expression with RT-qPCR. Cell extracts were analyzed for 14-3-3γ, phosphorylated IRF3 (pIRF3), total IRF3, influenza virus NP, and β-tubulin with immunoblotting.
Article Snippet: Human Embryonic Kidney 293T cell line (293T), adenocarcinomic human alveolar basal epithelial cell line (A549), and Madin Darby Canine Kidney (MDCK) cell line (obtained from the American Type Culture Collection (ATCC), Rockville, MD, USA) were cultivated in DMEM (Gibco, Waltham, MA, USA) supplemented with 10% fetal bovine serum (Gibco), 1% non-essential amino acids (Gibco), 1% penicillin-streptomycin (Gibco), and 1% L-glutamine (Gibco).
Techniques: Expressing, Infection, Transfection, Quantitative RT-PCR, Western Blot