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Lentiviral transduction for shRNA knockdown
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The shRNA plasmids for Adnp (TRCN0000081670; TRCN0000081671) and the GFP control (RHS4459) were purchased from Dharmacon (USA). To make lentivirus, shRNA plasmids and Trans-lenti shRNA packaging plasmids were co-transfected into H293T cells according to the kit manual (Open Biosystems, TLP4615). After determining the virus titer, mESCs were transduced at a multiplicity of infection of 5:1. Puromycin selection (1 μg/ml) for 4 days was applied to select cells with stable viral integration. qRT-PCR and western blotting were used to assess the knockdown of Adnp.
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