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. 2020 Aug 18;12(3):213–219. doi: 10.1007/s13238-020-00775-x

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© The Author(s) 2020

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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ZFP281 functions as a barrier in ESC-to-XEN differentiation. (A) A schematic plot of ESC-to-XEN differentiation in vitro. To avoid contamination of irradiated MEF feeders, RNAs were collected at P0 (passages 0), P1 and P3 (feeder-free) for qRT-PCR analysis. (B) Phase contrast microscope images of WT and Zfp281^−/− ESC-derived cXEN cells at P0-P3. White and red arrows indicate the XEN-like and ESC-like colonies, respectively. (C and D) qRT-PCR analysis of Zfp281 (C), pluripotency (Oct4, Nanog, Sox2) and PrE (Gata4, Gata6, Sox17) (D) transcripts in WT and Zfp281^−/− ESC-derived cXENs at P0, P1, and P3. (E) Immunostaining of NANOG and GATA6 at P1 and P3 in ESC-to-XEN differentiation. WT ESCs and XENs were used as positive controls for NANOG and GATA6 staining, respectively. (F) A volcano plot of differentially expressed genes (DEGs, fold-change > 2, T-test P-value < 0.01) between ESCs and XENs. DEGs highly expressed in ESC and XEN were defined as ESC-signature genes and XEN-signature genes, respectively. (G and H) Box plots for the expression of ESC-signature genes (G) and XEN-signature genes (H) in WT and Zfp281^−/− ESCs in cXEN differentiation. P-value was from a Mann-Whitney test

ZFP281 functions as a barrier in ESC-to-XEN differentiation. (A) A schematic plot of ESC-to-XEN differentiation in vitro. To avoid contamination of irradiated MEF feeders, RNAs were collected at P0 (passages 0), P1 and P3 (feeder-free) for qRT-PCR analysis. (B) Phase contrast microscope images of WT and Zfp281^−/− ESC-derived cXEN cells at P0-P3. White and red arrows indicate the XEN-like and ESC-like colonies, respectively. (C and D) qRT-PCR analysis of Zfp281 (C), pluripotency (Oct4, Nanog, Sox2) and PrE (Gata4, Gata6, Sox17) (D) transcripts in WT and Zfp281^−/− ESC-derived cXENs at P0, P1, and P3. (E) Immunostaining of NANOG and GATA6 at P1 and P3 in ESC-to-XEN differentiation. WT ESCs and XENs were used as positive controls for NANOG and GATA6 staining, respectively. (F) A volcano plot of differentially expressed genes (DEGs, fold-change > 2, T-test P-value < 0.01) between ESCs and XENs. DEGs highly expressed in ESC and XEN were defined as ESC-signature genes and XEN-signature genes, respectively. (G and H) Box plots for the expression of ESC-signature genes (G) and XEN-signature genes (H) in WT and Zfp281^−/− ESCs in cXEN differentiation. P-value was from a Mann-Whitney test