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. 2021 Jun 27;156(3):197–207. doi: 10.1007/s00418-021-02008-6

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© The Author(s) 2021

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/.

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Fig. 3 — Immunofluorescence analysis of human blastocysts. a Representative immunofluorescence images of human blastocysts analyzed for 5mC (magenta), 5hmC(cyan), and DNA signals (white). The major fraction of human blastocysts (17 of 24, upper panel) show stronger 5mC and 5hmC signals in the inner cell mass (ICM) compared to the trophectoderm; a smaller subset (7 of 24, lower panel) show weaker 5mC and 5hmC signals in the ICM compared to trophectoderm. The ICM is indicated by a yellow circle. Scale bar = 20 μm. b Number of cells per blastocyst with stronger or weaker 5mC and 5hmC signals in the ICM compared to trophectoderm. Each dot represents a single blastocyst. Statistically significant differences were calculated using two-tailed Student’s t test