HEK293T, HuH-7 and Caco-2 cells were maintained in DMEM supplemented with 10% FBS at 37°C in 5% CO2. Transient transfections were performed using Lipofectamine LTX (Invitrogen - Catalog #15338100) and GeneJuice (Novagen -Catalog #70967-3) according to manufacturer's instructions. Lentiviral infections of Caco-2 cells were performed as detailed in Langlois et al.72 (link) All transfections with multiple plasmids were performed using an equal amount of overall plasmid DNA. Differences in the amount of DNA from experimental constructs were filled with an empty construct.
FACS analysis was performed on a Accuri C6 cytometer (BD) after transfection/infection of the cells, at the times indicated in the figure legends.
Cell visualization was performed on an Eclipse TE2000-E confocal (Nikon) confocal microscope (Fig. 5), or on a Axio Observer Z1 (Zeiss) inverted microscope (Fig. S4). Transiently transfected cells were replated on coverslips after 24 h from transfection, and then fixed by 4% Formaldehyde and stained at 48 hr. Immunocgemistry for EGFP was carried out using a rabbit anti-GFP antibody (1:500; Molecular Probes – Catalog #A11122) and a AlexaFluor488 goat anti-rabbit IgG secondary antibody (1:500; Molecular Probes – Catalog #A11008).