Gene expression analysis of pluripotency and differentiation markers in differentiating ESCs. Normalized quantitative RT-PCR data from in vitro ESC differentiation assays. (A) Oct4 and (B) Nanog expression in ESCs, and in embryoid bodies (EBs) upon removal of LIF (day 2 [d2] liq, d5 liq). D8 MC time point is from EBs that were transferred to hematopoietic-inducing methylcellulose on d2 and grown for an additional 6 days. (C) Expanded gene expression analysis in d8 MC cultures of HAT^ptmut and p300^R/− cells. Gene expression levels in p300^R/− cells were all set to 1 and expression in HAT^ptmut cells is shown relative to this. (D) Dppa4 and (E) Gdf3 expression in d2 and d8 liquid EB cultures and after growth in cytokine-rich MC, d8 MC. Gene expression data were normalized to glyceraldehyde-3-phosphate dehydrogenase using the 2^−ΔCt method.³⁶  Bars indicate the average of triplicate samples; error bars indicate SD among triplicates. *Significant difference from p300^R/− (P < .001), calculated by Student t test. Graphs are representative of 2 to 4 independent experiments.