Validating Luciferase Transgene and IFN-β

For validation of luciferase transgene activity, we used the Luciferase Assay System kit as per the manufacturer’s protocol (Promega, Madison, WI, USA). Sample luminescence was read on a 1420 Multilabel Counter VICTOR3 (Perkin Elmer, Waltham, MA, USA). The luminescence results were normalized by protein concentration. Interferon-β detection was performed using the Verikine human IFN beta ELISA kit (PBL Assay Science, Piscataway, NJ, USA) and conditioned medium derived from transduced cell cultures. For p14ARF, we use immunofluorescence and Western blot as previously described (37 (link)–39 (link)), using anti-p14ARF antibody (Santa Cruz Biotechnology, Dallas, TX, USA). Immunofluorescence detection was obtained using EVOS FL Cell Imaging System (Thermo Fisher Scientific).