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. 2015 Oct 24;15:38. doi: 10.1186/s12861-015-0086-5

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© Freyer et al. 2015

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Fig. 3 — Gata6 ^{H2B-Venus/H2B-Venus} homozygous blastocysts exhibit a Gata6 null phenotype. a Immunofluorescence for GATA6 and NANOG protein in Gata6 ^+/+ (+/+; wild-type) Gata6 ^H2B-Venus/+ (Venus/+; heterozygous) and Gata6 ^{H2B-Venus/H2B-Venus} (Venus/Venus; homozygous) blastocysts at E3.5. Gata6 ^{H2B-Venus/H2B-Venus} mutants did not express GATA6 protein, and instead expressed NANOG in all ICM cells even though the reporter is transcriptionally active. Nuclei are stained with Hoechst (blue). b Quantification of cells with epiblast (EPI, NANOG+) versus PrE (PRE, GATA6+) identity at E3.5. Cells that did not adopt a clear identity were marked as double positive (DP) for both NANOG and GATA6. Gata6 ^{H2B-Venus/H2B-Venus} homozygotes did not specify PrE, and Gata6 ^H2B-Venus/+ heterozygotes had a relatively reduced number of cells with PrE identity and increased numbers of double positive cells with undecided identity. Gata6 ^{H2B-Venus/H2B-Venus} embryos with both alleles of the reporter, and effectively a Gata6 null, had higher levels of Venus expression compared to Gata6 ^H2B-Venus/+ embryos with only one reporter allele. This would suggest that GATA6 either does not regulate, or negatively feeds back on, Gata6 gene expression.