Lentiviral Transduction
U2OS WT cells were seeded at 300,000 cells in a 35 mm dish (50%
confluency) and cultured for 24 hours under standard culture conditions.
The number of viral particles were calculated according to the
multiplicity of infection (MOI), 5 MOI per U2OS cell, totaling 1.5 x
106 total transducing units needed. The appropriate
number of lentiviral particles (MATK-GFP-tagged), culture medium, and 4
µg/mL polybrene to the total volume of 500 µL was added to the 35 mm
dish and cultured overnight. The next day after transduction, the medium
containing lentiviral particles was removed and replaced with fresh
medium. 72 hours after transduction, a stable cell line was generated
using 4 µg/mL of puromycin (drug resistant marker).