Abstract
LIM kinases are involved in various cellular events such as migration, cycle, and differentiation, but whether they have a role in the specification of mammalian early endoderm remains unclear. In the present study, we found that depletion of LIMK2 severely inhibited the generation of definitive endoderm (DE) from human embryonic stem cells (hESCs) and promoted an early neuroectodermal fate. Upon the silencing of LIMK2 during the endodermal differentiation, the assembly of actin stress fibers was disturbed, and the phosphorylation of cofilin was decreased. In addition, knockdown of LIMK2 during DE differentiation also interfered the upregulation of epithelial-to-mesenchymal transition (EMT)-related genes and cell migration. Collectively, the results highlight that the serine/threonine kinase LIMK2, acting as a key regulator in actin remodeling, plays a critical role in endodermal lineage determination.
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Acknowledgements
We thank Xiaodong Shu from Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, for providing technical supports.
Funding
The work was supported by grants from Key Technologies Research and Development Program of China [grant number 2019YFA0111300], National Natural Science Foundation of China [grant numbers 31701183], Natural Science Foundation of Guangdong Province [grant number 2019A1515011324], Applied Basic Research Program of Sichuan Province [grant number 2021YJ0200], and Southwest Medical University [grant number 2020ZRQNA020].
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Editor: Tetsuji Okamoto
Supplementary Information
Supplementary Figure 1:
STR analysis of H1 cell line. (A) Match analysis. (B) Electropherogram of H1 cell line. (PNG 942 kb)
Supplementary Data 1:
siRNA sequences and primer sequences for RT-qPCR (XLSX 11 kb)
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He, Y., Zhang, L., He, Y. et al. Involvement of LIMK2 in actin cytoskeleton remodeling during the definitive endoderm differentiation. In Vitro Cell.Dev.Biol.-Animal 57, 493–500 (2021). https://doi.org/10.1007/s11626-021-00582-6
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DOI: https://doi.org/10.1007/s11626-021-00582-6