Human ifn α alpha 2a

The data described the expression of MxA and IFIT1 after IFN-α stimulation in a population of Huh7.5 cells. The experiments were done in the following way: Cells harboring a BAC (Bacterial Artificial Chromosome) containing the studied ISGs (mxa and ifit1) fused with the destabilized enhanced green fluorescent protein (deGFP) were seeded into a 6-well plate with 1.5x10⁵ cells per well. Next day, cells were treated in a time-dependent manner with IFN-α (8, 12, 16, 20 and 32 h) and harvested at the same time point. Cells were fixed with 2% paraformaldehyde to stop further protein expression for 10 min, washed three times with PBS and applied for flow cytometry (BD Accuri C6). Measuring GFP fluorescence intensity served as marker for ISG induction. Data was evaluated with the FlowJo v10 software package. Human IFN-α (Alpha 2a) was obtained from Pbl Assay Science with a reported specific activity of 3.91x10⁸ units/mg.

For cell experiments, cells were inoculated with HMPV strain TN/94-49 at an MOI of 1–10 in a 24-well plate. Mock-infected cells were inoculated with media or LLC-MK2 cell lysate, which had an equivalent effect on STAT1 and STAT2 protein levels and phosphorylation [30 ]. Then, 16–24 h post-infection, cells were treated with 1000 U/mL human IFNα (Alpha 2a) (PBL; Piscataway, NJ, USA) for human and primate cells, or 1000 U/mL murine IFNβ (PBL) for mouse cells for 30–40 min. After treatment, media were aspirated from the tissue culture dish and cells were lysed in RIPA buffer (ThermoFisher; Waltham, MA, USA) for western blotting or fixed in 4% paraformaldehyde for immunofluorescence.