Lentivirus production and transduction

Lentiviral production was performed using 293FT cells, which were co-transfected with pMD2.G (Addgene, #12259), psPAX2 (Addgene, #12260), and dTAG lentiviral plasmids using Lipofectamine 2000 (Thermo Fisher Scientific), according to manufacturer’s instructions. Viral particles were collected 48 and 72 hours after transfection, filtered through a 0.45 µM membrane, and concentrated using 1:3 Lenti-X Concentrator (Clontech), according to manufacturer’s instructions. Concentrated viral supernatants were applied to NIH/3T3 and 293FT cells in the presence of 4 µg/mL polybrene, and transduced cells were selected with 2 µg/mL puromycin. MV4;11 were transduced by spinoculation with concentrated virus for one hour at room temperature in the presence of 8 µg/mL polybrene, and transduced cells were selected with 2 µg/mL puromycin.