Fig. 1
From: Late stage definitive endodermal differentiation can be defined by Daf1 expression
Daf1+ DE cells are less proliferative than Daf1- DE cells. a Mouse ESCs (ESC) were differentiated first to the anterior primitive streak then into the definitive endoderm (DE). Our previously reported microarray analysis of Foxa2, Sox17 and Daf1 expression during DE differentiation are plotted in a time dependent manner. Daf1 expression increased in the DE but not in the APS. ESC; mouse embryonic stem cells, APS; anterior primitive streak, DE; definitive endoderm. b Schematic drawing of the experiment to analyze cell cycle phases of the sorted DE. ES cells were differentiated into DE, then dissociated and sorted for Daf1+/- cells. The sorted cells were immediately analyzed for cell cycle. c Real time-PCR analysis showed that Daf1 expression was enriched in Daf1+ DE. Y axis shows relative gene expression, with 1 equivalent to Daf1 expression level in Daf1 + DE cells. d, e Proliferative properties of Daf1 + DE and Daf1-DE cells were assayed. d Cell proliferation marker, pH3 (M phase), and PCNA (every cell phases except G0) was down-regulated in Daf1 + DE. Analyzed by western blot analysis. e Cell cycle analysis revealed that Daf1 + DE reside longer in the G0/G1 phase with a shorter S and M/G2 phases compared to that of the Daf1-DE cells. Raw data are shown in Additional file 2. Student’s two tailed t-test