Pluripotency-State-Dependent Role of Dax1 in Embryonic Stem Cells Self-Renewal

Figure 2

Establishment of Dax1KO-ESCs in 2i culture. (a) Schematic of the Cas9/sgRNA target site in the mouse Dax1 genomic locus. The sgRNA-targeting sequence is underlined, and the PAM sequences are marked in red. (b) Schematic diagram of lentivirus-based dual U6-sgRNA expression cassette constructs with the cassette of EFS-zeocin. (c) The expression of Dax1 and Actin was detected by Western blotting. (d) Dax1 cDNA was cloned into an expression vector carrying a CAG promoter and within the transcription unit linked to the puromycin resistance gene by an IRES. (e) Dax1-null colonies screened under LIF/serum and 2i conditions.

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