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. 2020 Jun 18;11:3101. doi: 10.1038/s41467-020-16885-3

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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. 4 — a, c, e Chromosome axis (SYCP3) and crossover markers (a) MLH1, (c) CDK2 or (e) HEI10 were detected by immunofluorescence in spreads of mid pachytene spermatocytes. c, e Bottom panels of each genotype show enlarged insets. Bars, 10 µm; in enlarged insets, 5 µm. b Quantification of MLH1 foci in mid/late pachytene spermatocytes. MLH1 was detected by two distinct antibodies raised in mouse and rabbit. Only the axis-associated foci detected by both antibodies were quantified. d Quantification of axis-associated interstitial CDK2 foci in mid pachytene spermatocytes. f Quantification of axis-associated HEI10 foci in mid/late pachytene spermatocytes. Strong foci of HEI10, which are thought to mark crossover-specific recombination complexes, were quantified separately from weak HEI10 foci along the axis. b, d, f n = numbers of analysed cells from two animals; medians (bars) were compared with Mann–Whitney U test, when possible, and are as follows in wild-type and Prr19^−/−: b 21 and 0 MLH1 foci, d 21 and 0 interstitial CDK2 foci (P = 2.59E-15), f 23 and 1 strong HEI10 foci (P = 7.26E-15), 19 and 75 weak HEI10 foci (P = 1.25E-14), respectively. **** indicates P < 0.0001. g DNA was labelled with Hoechst 33342 in spread diakinesis/metaphase I spermatocytes. Arrow indicates univalent in Prr19^−/− cell. Bars, 20 µm. h Quantification of bivalents in spread diakinesis/metaphase I spermatocytes from mice of indicated genotypes. n = numbers of analysed cells from three (wild-type and Prr19^−/−) or two (Mlh1^Lisk/Lisk and Prr19^−/− Mlh1^Lisk/Lisk) mice; medians (bars) are indicated. Median bivalent numbers are as follows: 20 in wild-type, 3 in Prr19^−/−, 2 in Mlh1^Lisk/Lisk and Prr19^−/− Mlh1^Lisk/Lisk. Results of Mann–Whitney U test are shown. **** indicates P < 2.2E-16 between wild-type and Prr19^−/−, ns indicates no significance between Prr19^−/− and Mlh1^Lisk/Lisk (P = 0.0636), between Prr19^−/− and Prr19^−/− Mlh1^Lisk/Lisk (P = 0.0763) and between Mlh1^Lisk/Lisk and Prr19^−/− Mlh1^Lisk/Lisk (P = 0.9966). Source data are provided as a Source Data file.