Complementary DNA transfection was carried out using Lipofectamine 3000 (Life Technologies). To conduct shRNA-mediated gene silencing, pLKO.1 puro (Addgene, 8453) constructs containing 19–base pair gene-specific shRNAs were first packed into virus by cotransfecting 293T cells with psPAX2 and pMD2.G. Twenty-four hours after transfection, the lentivirus was collected, filtered through a 0.44-μm filter, and used to infect target cells. Forty-eight hours after infection, the virus-containing medium was removed and the target cells were selected using puromycin at the concentration of 10 μg/ml. Knockdown efficiency was confirmed by Western blotting.

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