The corresponding three shRNAs for Bach1 or control shRNA was cloned into the pLKO.1 constructs (no. 8453; Addgene, Watertown, MA). Lentiviral particles of three Bach1-shRNAs or control shRNA was generated by transfecting HEK293T cells with a pLKO.1 shRNA vector and packing vectors psPAX2 and pMD2.G. hESCs were transduced with the concentrated lentivirus for 48 hours; then, the cells were isolated with puromycin (2 μg/ml) selection, and clones were chosen for use in subsequent experiments. The sequences of Bach1 shRNAs are shown in table S2.4.

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