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Previously, we have shown that exposure of mouse embryonic stem cells (mESCs) to high concentrations of diethylenetriamine nitric oxide adduct (DETA-NO) triggers differentiation events and promotes the expression of Pdx1. Here we report evidence that Pdx1 expression is associated with release of polycomb repressive complex 2 (PRC2) and P300 from its promoter region. These events are accompanied by epigenetic changes in bivalent markers of histones trimethylated histone H3 lysine 27 (H3K27me3) and H3K4me3, site-specific changes in DNA methylation, and no change in H3 acetylation. On the basis of these findings, we developed a protocol to differentiate mESCs toward insulin-producing cells consisting of sequential exposure to DETA-NO, valproic acid, and P300 inhibitor (C646) to enhance Pdx1 expression and a final maturation step of culture in suspension to form cell aggregates. This small molecule-based protocol succeeds in obtaining cells that express pancreatic \u03b2-cell markers such as PDX1, INS1, GCK, and GLUT2 and respond in vitro to high glucose and KCl.<\/jats:p>","DOI":"10.3727\/096368916x691178","type":"journal-article","created":{"date-parts":[[2016,3,15]],"date-time":"2016-03-15T03:04:14Z","timestamp":1458011054000},"page":"1879-1892","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":17,"title":["Differentiation of Mouse Embryonic Stem Cells toward Functional Pancreatic \u03b2-Cell Surrogates through Epigenetic Regulation of\n                    <i>Pdx1<\/i>\n                    by Nitric Oxide"],"prefix":"10.1177","volume":"25","author":[{"given":"Carmen","family":"Salguero-Aranda","sequence":"first","affiliation":[{"name":"Andalusian Center for Molecular Biology and Regenerative Medicine, Seville, Spain"},{"name":"Center for Network Research in Diabetes and Metabolic Diseases 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