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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. 7 — a, b, e Immunofluorescence staining of surface-spread mid/late pachytene spermatocytes. b Bottom panels show enlarged insets. a, b, e Bars, 10 µm, or (bottom panel, b) 5 µm. c Quantification of co-localisation between CNTD1 and PRR19 foci on the chromosome axis in mid/late pachytene wild-type spermatocytes, medians (bars) are 96.4% (blue data set) and 95.7% (red data set). d Quantification of axis-associated CNTD1 focus numbers in wild-type and Prr19^−/− mid/late pachytene spermatocytes. Medians (bars) are 24 and 3 in Prr19^+/+ and Prr19^−/−, respectively. Mann–Whitney U test, P < 2.2E-16 (****). c, d n = numbers of analysed cells from two animals. f, h Immunoblots of immunoprecipitation experiments from testis extracts of (f) 13-days-old or (h) adult mice. Arrowheads mark bands of indicated proteins. Asterisks mark unspecific protein bands. Molecular weights are indicated. f, upper panel: PRR19 was detected in immunoprecipitates (IP) of guinea pig anti-PRR19 antibodies to assess PRR19 levels without interference from unspecific immunoblot signals that are present in total extracts. f, two middle panels: CNTD1 in anti-PRR19 IPs and corresponding input samples. f, bottom panel, anti-α-tubulin used as loading control. h Images show input for immunoprecipitation, immunoprecipitates (IP) of guinea pig anti-PRR19 (Gp-PRR19), anti-CNTD1 (Gp-CNTD1) and non-specific (Gp–IgG) antibodies. g Immunoblot signals of PRR19 (from anti-PRR19 IPs) or CNTD1 (from testis extracts) in the indicated genotypes were normalised against corresponding immunoblot signals of wild-type littermate controls. Two-tailed one-sample t test calculated significance of difference between the wild-type value of one and means of six experiments (bars) for PRR19, 0.05 (P = 3.63E-09), 0.19 (P = 2.17E-06) and 0.19 (P = 3.52E-07), and CNTD1, 0.34 (P = 6.82E-05), 0.03 (P = 5.42E-09) and 0.33 (P = 9.27E-05), in Prr19^−/−, Cntd1^−/− and Cntd1^Q/Q samples, respectively. **** indicate P < 0.0001. i Yeast two-hybrid interaction assays between indicated proteins. CNTD1Q refers to the protein product of Cntd1^Q allele (see Supplementary Fig. 3). Yeast cultures are shown after 2 or 3 days of growth on dropout plates. For negative control, proteins of interest were tested in transformations where either the Gal4-binding domain (Gal4-BD) or the Gal4-activation domain (Gal4-AD) vectors were empty. Source data are provided as a Source Data file.