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. 2023 Aug 22;30(10):2293–2308. doi: 10.1038/s41418-023-01207-z

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© The Author(s) 2023

Open Access This 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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Fig. 6 — A Co-Immunoprecipitation (co-IP) assay between Hnrnpk and Ctcf at E11.5 in WT limb bud after treatment with PBS, DNase I, RNase A, or both. B Subcellular location of Hnrnpk and Ctcf at E11.5 in primary WT limb bud cell. The white arrows indicated the overlapped Hnrnpk and Ctcf signals in the nucleus. The white dotted line indicated the margin of nucleus. Scale bar: 10 μm. C The Hnrnpk- and Ctcf- binding peaks at E11.5 in WT limb buds and their overlap. The ratio indicated the proportion of overlapped peaks in the total peaks of Hnrnpk. D, E The binding profile of Ctcf centered in Hnrnpk binding peaks. F Protein level of Ctcf at E11.5 in WT and CKO limb buds. β-Actin was used as the loading control. G Hi-C contact map (upper) and genome browser tracks of Hnrnpk and Ctcf CUT&RUN-seq data (bottom) at Shh-TAD. The black arrows indicated the decreased binding strength of Ctcf at E11.5 in CKO limb bud. The yellow lines indicated the boundaries at both sides of Shh-TAD. H The Ctcf binding peaks centered in CTCF-motif at E11.5 in WT and CKO limb buds. I A proposed model illustrated the role of Hnrnpk as a transcription activator and insulator protein collaborator, which regulated the transcription of key regulatory genes during embryonic limb bud development.