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. 2018 Dec 15;7(12):bio036517. doi: 10.1242/bio.036517

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© 2018. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 1. — Generation of Gata4^H2B-GFP/+ targeted mouse model. (A) The cassette LoxP-H2B-GFP-4XPloyA-LoxP-FRT-Neo-FRT-3XFLAG was inserted into the start codon of the mouse Gata4 locus through homologous recombination. The Neo cassette was flanked by two FRT sites and removed by crossing to Flippase mice to generate the Gata4^H2B-GFP allele. Cre excision of H2B-GFP generates GATA4-FLAG N-terminal protein fusion Gata4^FLAG allele. (B) The targeted ES cells were confirmed by long-range PCR amplification of two fragments using primers P1 and P2, P3 and P4, respectively. Primers P1 and P2 produced a 4.3 kb band. Primers P3 and P4 produced a 4.9 kb band. (C) Western blot of individual E7.5 embryo littermates from a Gata4^FLAG/+×wild-type mating. The anti-FLAG antibody cross reacts with GATA4-FLAG in Gata4^FLAG/+ embryos. Total protein lysis of an ear from a Gata4^FLAG/+ adult male was used as a positive control.