FU-GFP-IRES-PURO-W lentivirus constructs were used for pMGL transduction. VSVG-coated lentiviruses were generated in HEK293 cells as previously described 50. Sub-confluent HEK293 cells were transfected using X-tremeGENE 9 (Roche), with a mixture of lentiviral construct and second-generation packaging plasmids. Culture medium was changed 12 hours after transfection and collected 96 hours later. Virus-containing medium was filtered through 0.45μm filter and concentrated by ultracentrifugation (23krpm, 90′, 4°C). pMGLs cultured in MGdM were transduced at an MOI of 10 (titer assessed by p24 ELISA). Medium was replaced after 12 hours, and cells were left to recover. Expression was evaluated by fluorescence microscopy without additional selection.
