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Cells were plated in 6-well plates at 7 10 5 cells per well and transfected with 2.5 g of the appropriate TCR or CAR construct in the retroviral vector pMP71 using Lipofectamine 3000 (Life Technologies, Invitrogen)

Western blotting After treatment with bortezomib (4 nM), UroA (15 M), their combination, or vehicle (as a control), cells were lysed and cleared by centrifugation (14,000 g , 4 C, 5 min), and the protein concentration was determined with the Quick Start Bradford 1 Dye Reagent (Bio-Rad Laboratories, Hercules, CA, USA) on the Epoch microplate reader

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In that respect, IM8 comes in at a sort of bargain, although its up-front price tag remains beyond reach for most people