Figure 1.

Differentiation of Mixl1-early, Mixl1-intermediate and Mixl1-late EpiSCs. (a) Level of expression of Mixl1 (PS marker), Sox17 and Foxa2 (definitive endoderm marker) in day-4 EBs cultured under seven culture protocols. Gene expression level is presented as the negative value of the relative difference in threshold cycles (−ΔΔCT; normalized against Actb) relative to day 0 value and is the mean of triplicate cultures. Positive value, upregulation; negative value, downregulation. (b) Endoderm cells co-expressing Foxa2 and Sox17 (arrowheads) in day-4 EBs, visualized by immunofluorescence of Foxa2 (red) and Sox17 (green). (Online version in colour.)