Generation of shRNA-containing lentivirus

One million Hek293 FT cells were plated per well in a six well culture plate (Corning) and transfected 2 h post-seeding with a 4–2-1 ratio of Plasmid: gagpol:vsvg, PEI transfection reagent (Sigma) in 100 µl serum-free DMEM (LifeTech). Cells were transfected overnight, and media replaced the following morning with 2 ml DMEM, 10%FCS, and 5% PenStrep. Cells were incubated for a further 24 h before lentiviral-containing media were removed and filtered through a 0.45 µm filter, lentiviral-containing media were then stored at −80°C until use. shRNA vector details are provided in ^{Table 3}.