Huh7.5 cells (a kind gift of Prof. R. Bartenschlager, Heidelberg University, Germany, with the authorization of Apath LLC, NY) (52 (link)) were cultivated in DMEM high glucose (Gibco) supplemented with 10% calf serum. For the growth assays, cells were seeded in 6–96-well plates (cell culture treated, Corning, New York, USA). After the indicated time, the cells were collected and lysed with CellTiter-Glo Luminescent 3D Cell Viability Assay reagent (Promega, Mannheim, Germany). Caspase activity was evaluated using the ApoTox Triplex Assay (Promega). Luminescence was analyzed by the Spark Microplate Reader (Tecan, Männedorf, Switzerland).
Huh7.5 transduction was carried out with lentiviral vectors, as previously described (34 (link)), with third-generation lentiviral vectors to transduce simultaneously both reporter and miR34a.
Sofosbuvir, velpatasvir, and grazoprevir (Selleckchem, Boston, USA) were resuspended as indicated by the vendor and used at the concentration of 10 nM; the absence of the virus was assessed as described below.
Huh7.5 transduction was carried out with lentiviral vectors, as previously described (34 (link)), with third-generation lentiviral vectors to transduce simultaneously both reporter and miR34a.
Sofosbuvir, velpatasvir, and grazoprevir (Selleckchem, Boston, USA) were resuspended as indicated by the vendor and used at the concentration of 10 nM; the absence of the virus was assessed as described below.