Fig. 1

Time course analysis of expression of genes associated with pluripotency and early differentiation of human embryonic stem cells (hESCs) in response to inhibition of glycogen synthase kinase-3 (GSK-3). a Schematic representation of differentiation of hESCs with or without the inhibition of GSK-3 using CHIR99021 (±GSKi). Real-time RT-PCR analysis of markers associated with pluripotency and neuroectoderm (b), primitive streak (PS)/mesendoderm, epithelial–mesenchymal transition (EMT), anterior PS/endoderm, and mesodermal subsets (c) after differentiation of hESCs with (grey bars) or without (black bars) GSKi. For all gene expression plots, expression levels were normalized to corresponding β-ACTIN values and are shown relative to undifferentiated hESCs. The expression levels of SOX2, OCT4, NANOG, PAX6, SOX1 and VEGFR2 were log-normalized to reveal the amount of downregulation in relation to undifferentiated hESCs. Error bars show standard deviations; n ≥ 3. *p < 0.05, **p < 0.01, versus without GSKi. d Confocal immunofluorescence micrographs show expression of OCT4 and BRACHYURY after differentiation with or without GSKi. Scale bars = 200 μm. DMSO Dimethyl sulfoxide,