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siRNA transfection
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Cells were seeded at a density of 3×105 cells/well in 6-well plates. The cells were transfected with siRNA using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) following the manufacturer's instructions. siRNA-control and siRNA-AKT (Santa Cruz Biotech, Dallas, TX, CA) were used. Western blot analysis was used to verify the gene silencing efficiency. After 48 h, cells were pretreated with gefitinib and LC for cell growth inhibition analysis.
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