The cells were transfected with plasmids containing the gene of interest (Sangon Biotech (Shanghai) Co., Ltd.) using Lipofectamine 2000 (Invitrogen, 11668-019) transfection reagent. The cells were seeded on 6-well plates in 2% FBS DMEM/1640 medium. Once the cells attached to the plate, they were transfected with the plasmids (final DNA used per well: 2,500 ng) using 10 µl of Lipofectamine 2000 per well following the manufacturer's protocols. The transfection reagents were removed after 12 h, and the cells were treated as indicated in each experiment. Western blotting and reverse transcription polymerase chain reaction (RT-PCR) were used to verify the transfection efficiency.

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