Human ifn α 2b

Cell lines or ovarian patient ascites cells were cultured in ACM overnight. The next day the cells were treated with 1μM Jaki (EMD Millipore) for 3 hours before receiving 200 U/mL human IFN-α 2b (Sigma Aldrich). After 3 hours of IFN-α treatment, the cells were infected with VSVΔ51.

Hepatocellular carcinoma HepG2 cell line was procured from American type culture collection (ATCC). We propagated HepG2 cells in Dulbecco’s modified Eagle’s medium (GBICO) supplemented with 10% fetal bovine serum (GBICO) in a humidified incubator containing 5% CO₂ at 37 °C. Human IFN-α2b (Sigma-Aldrich) was diluted in serum-free medium.

Subconfluent A549 cells were treated with 1,000 U/mL human IFN-α2b (SRP4595, Sigma-Aldrich) 24 h prior to infection with HAdV wt. The cells were harvested at 48 hpi for mRNA isolation, cDNA transcription, and a qPCR analysis for Apobec3A transcription, while a control setup was treated and infected equally to harvest the interferon secreted supernatant at 72 h post treatment. This time point was chosen to exclude potentially remaining interferon from the initial treatment. The dilution rows of supernatant were added for 6 h to the reporter cell line HL116, which expressed a stably integrated luciferase gene under the control of the interferon-responsive promoter region 6 to 16 (36 (link)). Dual luciferase reporter assays were performed to measure the interferon secretion, and the median values were calculated.