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. Author manuscript; available in PMC: 2013 Feb 3.
Published in final edited form as: Stem Cells. 2009 Dec;27(12):2884–2895. doi: 10.1002/stem.203

Figure 5. Mesendoderm and mesoderm marker genes are activated in differentiating i-Mixl1 ES cells.

Figure 5

(A) General experimental protocol for analysis of gene expression patterns of differentiating i-Mixl1 ES cell monolayer cultures using QRT-PCR. i-Mixl1 ES cells were allowed to differentiate in the LIF-free medium in the absence (−) or presence (+) of 0.4 μg/ml of DOX. Total cellular RNA was isolated at days 1 through 5 of culture and reverse transcribed to cDNA for QRT-PCR analysis. (B) The expression of endogenous Mixl1 reached a peak at day 3 and declined at day 5 of culture in uninduced cells (−DOX). Induction of Mixl1 (+DOX) accelerated endogenous Mixl1 expression. (C) The expression of Gsc was peaked at day 3 and declined at day 4 of culture in uninduced cells (−DOX). Induction of Mixl1 expression (+DOX) resulted in a premature activation of Gsc as early as day 1. (D) The expression of the Brachyury (T) gene was activated at days 3–4 and declined at day 5 of culture in uninduced cells (−DOX). Induction of Mixl1 (+DOX) accelerated T expression at day 2 of culture.