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. 2023 Apr 24;24:92. doi: 10.1186/s13059-023-02925-w

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© The Author(s) 2023

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. 5 — WNT signaling pathway acts as downstream of HIDEN/IMP1. a Venn diagram indicates the overlapped genes of differentially expressed PCGs upon HIDEN knockout in DE cells and IMP1-enriched targets identified by IMP1 RIP-seq in DE cells. b GO analysis of the overlapped genes from a. c, d HIDEN knockout led to reduced active β-catenin and unaltered total β-catenin level compared to wildtype after two days’ DE differentiation, as shown by Western blot (c) and statistical results (d) (n = 6). e The protein level of β-catenin in nuclear fraction of WT or HIDEN-KO DE cells (n = 3). f The TCF-luciferase activity in 293 T when transfected with HIDEN, antisense control or empty vector (n = 3). Cells treated with 1 μM CHIR-99021 were used as positive controls. g-j Flow cytometric analysis of SOX17-positive cells (g), the presentative endoderm genes expression revealed by RT-qPCR (h) and immunostaining (i-j) in wildtype or HIDEN-KO cells after manipulating WNT signaling through small molecular inhibitors during DE differentiation (n = 3). WNT signaling activator CHIR-99021, with different concentrations (0.5 μM, 1 μM and 2 μM) at (g) and 1 μM at (h-j), and WNT signaling inhibitor WNT-C59 (1 μM) were used. NT indicated for non-treated group. i Scale bar = 50 μm. j Quantitative results of SOX17- and FOXA2-positive cells were shown (n = 6)