MP incorporation modulates gene expression patterns. The gene expression of (A) primitive streak markers (mixl1 and brachyury-T), (B) EMT and early mesendodermal markers (E-cadherin N-cadherin, Slug, mmp2, mesp1, and gata4), and (C) lineage-specific markers (nanog, pax6, foxA2, and flk1) in aggregates at days 4, 7, or 10 of differentiation was analyzed using real-time PCR and normalized to housekeeping gene s18. Relative fold changes over ESC gene expression levels were calculated for all samples. * P<0.05 (n ≥ 4).