Gene expression of early germ-layer specific markers at stage 1 by qRT-PCR.

<p>Cells were induced to differentiate as EB in the presence of 100 ng/ml activin A as indicated in Fig. 1. After 3 or 5 days (stage 1), cultures were harvested and subjected to qRT-PCR analysis for the indicated early germ-layer (A–C) and foregut/pancreatic (D) markers. Histograms show the relative expression levels normalized to the loading control Hprt. Error bars indicate the standard deviations of 4 experiments. (d), days; T, treated cells; NT, non-treated cells. In A–C, p is calculated as compared to D1 and in D, as compared to T3. In D, the expression was analyzed from day 3 onwards as some of the markers were undetectable at day 1.