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. 2022 Nov 4;12:181. doi: 10.1186/s13578-022-00917-0

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© The Author(s) 2022

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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 3 — LEFTY-NODAL Signaling Pathway is Essential for hPGCLCs Differentiation. A Scheme of hPGC differentiation through iMeLCs in vitro; SB431524: NODAL signaling inhibitor; ACTA (Activin A): NODAL signaling stimulator; B Bright field (BF) and fluorescence images of day4 embryoids stimulated by cytokines and with or without SB431542 in iMeLCs or hPGCLCs induction. Scale bar = 100 μm; C FACS patterns show the induction efficiency of hPGCs with or without SB431542 at iMeLCs or hPGCLCs induction; D FACS analysis of WT and TKO hESCs in day4 hPGCLCs induction with cytokines and different concentration of Activin A; E FACS analysis of WT and TKO hESCs in day4 hPGCLCs induction with different concentration of Activin A and without cytokines; F mRNA expression was assayed in day4 embryoids treated with different concentration of Activin A without cytokines using qRT-PCR assay, the WT day4 hPGCLCs were used as positive control; n = 3 independent experiments. Data are presented as means ± s.d