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. 2023 May 29;14:3101. doi: 10.1038/s41467-023-38770-5

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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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a, b, Immunofluorescence for Lamin-A/C in fixed mouse embryos (a) and live-imaging of mRuby-Lamin-A (b) at different developmental stages. Images show thin confocal sections to illustrate inner-outer differences. Lamin-A L:N plotted against distance to embryo center of mass. For (a) n = 6 for 16-cell inner, n = 40 for 16-cell outer, n = 21 for blastocyst ICM, n = 33 for blastocyst trophectoderm. For (b) n = 29 for 16-cell inner, n = 48 for 16-cell outer, n = 18 for blastocyst ICM, n = 32 for blastocyst trophectoderm. ****P < 0.0001, Mann-Whitney U test. c, Live-imaging and quantification of mRuby-Lamin-A in embryos treated with H-1152 or blebbistatin show reduced mRuby-Lamin-A L:N. Conversely, embryos expressing the constitutively active RhoA mutant Q63L show increased mRuby-Lamin-A L:N. n = 11 for control outer, n = 5 for control inner, n = 11 for H-1152 outer, n = 6 for H-1152 inner, n = 10 for blebbistatin outer, n = 6 for blebbistatin inner, n = 13 for RhoA Q63L outer and n = 6 for RhoA Q63L inner. ***P < 0.0007, *P = 0.0343 for control outer vs blebbistatin outer, *P = 0.0410 for control outer vs RhoA Q63L outer. *P = 0.0418 control inner vs RhoA Q63L inner. Mann-Whitney U test. d, Human blastocyst immunostained for Lamin-A/C shows lower Lamin-A L:N in the ICM versus trophectoderm (TE). Quantification shows the relationship between Lamin-A/C L:N and distance to embryo center of mass. n = 19 for ICM, n = 108 for trophectoderm (two human blastocysts were analyzed and therefore statistical analysis was not performed). e, Human blastocysts treated with ROCK inhibitor H-1152 have reduced Lamin-A/C L:N (one human blastocyst per group, trophectoderm cells n = 199 for control, n = 95 for H-1152. All statistical tests are two-tailed. Bars in dot plots represent median and interquartile range. Scale bars, 10 µm. Source data are provided as a Source Data file.