Fig. 6

ZNF207 works upstream of OTX2 in hESCs to govern specification to ectoderm. a Immunofluorescence staining of control and ZNF207 KD hESCs for lineage markers. Scale bars represent 50 µm. b Counts and percentage of cells that were stained positive for each lineage. c KD of ZNF207 disrupted pluripotency of hESCs. HESCs have equal potential to differentiate into any of the three lineages, while KD of ZNF207 tilts the potential to endoderm and mesoderm. The potential to differentiate into ectoderm is significantly impaired by KD of ZNF207. d Immunofluorescence staining of control, ZNF207 KD and ZNF207 KD, OTX2 OE hESCs for ectodermal proteins. Top: Scale bars represent 100 µm; Bottom: Scale bars represent 25 µm. e Counts of cells that were stained positive for ectodermal proteins. f ZNF207-centered transcriptional network to regulate self-renewal and pluripotency in hESCs. ZNF207 works with master transcription factors and co-factors, such as OCT4, SOX2, and P300, to form the core transcriptional network in hESCs. They bind together at pluripotency, development, and cell cycle-related genes to govern self-renewal and pluripotency. OTX2 is targeted and regulated by ZNF207 to ensure the developmental potential towards ectoderm. Once ZNF207 is knocked down, the expression of OCT4, SOX2, and P300 is reduced. The level of cell cycle-related genes has been changed, which leads to reduced ability of self-renewal. The expression of OTX2, which is downstream of ZNF207, is also downregulated, which impedes the differentiation potential towards ectoderm. In the meantime, the potential to endoderm and mesoderm is enhanced. Data are presented as the mean ± SEM and are derived from three independent experiments