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. 2021 Sep 30;22(19):10620. doi: 10.3390/ijms221910620

Figure 1.

Figure 1

Ol-Nanog is functionally equivalent to human NANOG. (a) Nanog transcription factor binds to some genes at their promoters in medaka as it does in mESC. ChIP assay using the Ol-Nanog antibody. To determine the promoter sequences of the genes the conservation analysis was made using the genome sequences of Medaka, Stickleback, Tetraodon, Fugu, and Zebrafish. ChIP was performed with antibodies for Ol-Nanog, and Ol-Ptc1 as negative control. PCR reactions were performed before (pre-IP; positive control) and after ChIP. Abbreviations: Anti-Ptc1, Anti-Patched1; Pre-IP, pre-immunoprecipitation, P: regulatory region of the gene, N: sequence outside the regulatory region of the gene. (b–e) Ol-Nanog and h-NANOG rescue the abnormal phenotype provoked by MO-Nanog. (b–d) Pictures showing stage 31 embryos after injection in one cell stage of MO-Std as a MO control (b), MO-Nanog (c) and MO-Nanog+h-NANOG mRNA (d). While 100% of embryos injected with MO-Nanog that survive to stage 31 had small size and abnormal development of the head and trunk structures, 29.4% of embryos injected with MO-Nanog + h-NANOG showed normal size and head and trunk structures, when compared to embryos injected with MO-Std. Scale bars: 200 μm.