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. 2020 Feb 11;30(3):256–268. doi: 10.1038/s41422-020-0281-1

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© The Author(s) 2020

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 Numbers of PRDM9-mediated H3K4me3 peaks in leptotene (L), early-zygotene (eZ), mid-zygotene (mZ), late-zygotene (lZ), early 1-pachytene (e1P), early 2-pachytene (e2P), and mid-pachytene (mP) spermatocytes. b Profile of the averaged H3K4me3 tag density on early-forming hotspot-associated H3K4me3 peaks in leptotene, mid-zygotene, late-zygotene, and early 1-pachytene. H3K4me3 tag density was calculated using H3K4me3 ChIP-seq read coverage with 50-bp resolution. c Heatmaps of H3K4me3 (left) and H3K4me1 (right) normalized tag density on hotspot-associated H3K4me3 peaks in premeiotic and meiotic cells. Each row represents a hotspot-associated H3K4me3 peak of ±5 kb around the peak center and ranked based on SPO11-oligo density from the highest to the lowest. d Histone Methyltransferase Assay using recombinant Prdm9 and oligonucleosomes. The percentage of each methylation state on H3K4 site was quantified by histone mass spectrometry. The blank box represents no enzyme control. Undiff undifferentiated spermatogonia, A1 type A1 spermatogonia, B type B spermatogonia, mpL mid-preleptotene spermatocytes, L leptotene spermatocytes, mZ mid-zygotene spermatocytes, lZ late-zygotene spermatocytes, e1P early 1-pachytene spermatocytes, mP mid-pachytene spermatocytes, D diplotene spermatocytes.