Generation of Adnp−/− ESCs.a Cartoon depicting the gRNA target sites at exon 4 of mouse Adnp gene. b Genotyping showing the mutant alleles. c Western blot analysis of ADNP levels in control, shRNA knockdown, and Adnp−/− ESCs. d Representative image showing morphology of control and early passaged Adnp−/− ESCs. e The mRNA expression of representative pluripotency-related, mesodermal, neuroectodermal, endodermal genes in control, and early passaged Adnp−/− ESCs (n = 3 per group). f Representative image showing morphology of embryoid bodies (EBs) at indicated time points. g Heat-map analysis of DEGs of the indicated lineage-specific genes for control and Adnp−/− ESC-derived day 6 EBs, based on three RNA-seq replicates. h qPCR analysis showing the dynamic expression of the indicated genes during EB formation of control and Adnp−/− ESCs. qRT-PCR was based on three biologically independent experiments (n = 3 per group). Data are presented as mean values ± SEM in (e, h). p values by two-tailed unpaired t-test are shown in (e, h). ns: not significant. Source data are provided as a Source Data file. Experiments were repeated at least two times in (c, d, f), and similar results were obtained.
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