Lentiviruses were prepared according to the manufacturer’s protocol (GIPZ Lentiviral shRNA, Dharmacon). CT26, Treg, and CD8+ cells were infected by β-cat shRNA lentiviruses according to the manufacturer’s protocol using the Dharmacon GIPZ Lentiviral shRNA Kit (GE Healthcare). The nontargeting (NT) lentiviral shRNA construct expresses an shRNA sequence with no substantial homology to any mammalian transcript, providing a negative control. Cells were infected with NT, β-cat shRNA1, and β-cat shRNA2 lentiviruses as described in our previous studies, and β-cat protein level was evaluated by immunoblotting (5).

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