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. 2019 Apr 23;10:1884. doi: 10.1038/s41467-019-09713-w

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© The Author(s) 2019

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. 1 — Phenotypic description of mice carrying the Dnmt3a^D329A allele. a Schematic representation of the DNMT3A protein isoforms and location of the base substitution, coloured red, introduced to generate the D329A mutation. NCBI IDs: CCDS36397.1, CCDS25784.1. Known domains are indicated by coloured boxes, based on the PROSITE database; numbers indicate amino acid residues. b Breeding scheme used to generate offspring of four different genotypes. +; wild-type allele; fl: Dnmt3a containing loxP sites surrounding exon 18; Δ: loxP sites after recombination leading to deletion of exon 18; Zp3-Cre: oocyte-specific Zp3 promoter-driven Cre recombinase^21,22; D329A: a Dnmt3a allele point missense mutation in exon 8. c Growth curves indicating change in body weight of male and female mice of different genotypes over the indicated post-natal weeks. Mixed model ANOVA (KR-method) shows overall significant change dependent on genotype and time in both sexes (p < 0.001, df(males) = 15,229.48, df(females) = 15,142.15), F = 0). Subsequent pairwise Bonferroni-adjusted t-test comparisons show that Dnmt3a^Δ/D329A males become significantly underweight at week 4 (p < 0.001), and Dnmt3a^+/D329A at week 6 (p < 0.001). Dnmt3a^Δ/D392A females become significantly underweight starting from week 4 (p = 0.002), with the exception of week 8 (p = 0.095). Dnmt3a^+/D329A females do not show significant change. n values are given in Supplementary Table 3. Error bars indicate standard error of the mean. Raw data are provided in Source Data. d Volcano plot showing gene expression fold change and its significance between Dnmt3a^+/+ and Dnmt3a^Δ/D329A adult (14-week) hypothalamus. Differentially expressed genes were determined using DEseq (p < 0.01, Benjamini–Hochberg multiple comparisons correction). n(+/+) = 5, n(Δ/D392A) = 4. Selected genes showing the most significant and highest fold change are named. e Gene ontology (GO) analysis of differentially expressed genes; only significant terms are shown. Benjamini–Hochberg corrected p-value < 0.1 was used as a threshold