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. 2019 Nov 20;2:422. doi: 10.1038/s42003-019-0663-4

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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. 2 — MamoAF directly binds vas and epigenetically activates vas together with CBP. a vas-A element, which contains a Mamo-binding consensus sequence (red), in the first intron of vas. b EMSA analyses of Zn-finger domains of Mamo protein (MZD) using vas-A and vas-AΔ probes. vas-AΔ probe lacks a Mamo-binding consensus sequence. c ChIP-qPCR assays were performed with MamoAF-OE embryos and FLAG antibody. The precipitated DNA was detected by qPCR with primers targeting to vas-A. Error bars are SD of the mean. **P < 0.01 (two-tailed Student’s t test). d Vas mRNA in situ hybridisation of wild type, vas^d10-3 / + and vas^d10-3 / vas^d10-3 embryos at stage 10. The Mamo-binding consensus sequence (red box) is deleted from vas-A element in vas^d10-3 / vas^d10-3embryo. Scale bar: 50 μm. e ChIP-qPCR assays were performed with MamoAF-OE embryos and H3K27ac antibody. The precipitated DNA was detected by qPCR with primers targeting to vas-A. Error bars are SD of the mean. *P < 0.05 (one-tailed Student’s t test). f qRT-PCR analyses of Vas mRNA in wild type, MamoAF, CBP and both CBP and MamoAF overexpressed embryos. Error bars are SD of the mean. *P < 0.05, **P < 0.01 (two-tailed Student’s t test). g Vas mRNA in situ hybridisation of wild type and CBP^M- embryo derived from nej^0.3 mutant germline clones at stage 9. White arrowheads indicate PGCs expressing Vas mRNA. Scale bar: 50 μm. h Percentages of embryos expressing Vas mRNA in wild type and CBP^M- embryos at stage 9. *P < 0.01 (Fisher’s exact test)