After 35 days of differentiation, we observed expression of TUJ1 and MAP2AB in the EBs, indicating the derivation of neurons using the differentiation paradigm (Figure 1B). Strikingly, we detected a significant expression of PROX1 in the EBs (∼32%), a marker specific to DG granule neurons in the hippocampus (Hodge et al., 2012; Iwano et al., 2012), compared to the untreated group (∼4.7%) (Figures 1B and 1C). We further confirmed this observation using quantitative RT-PCR (qRT-PCR), which showed a remarkable increase in PROX1 expression (Figure 1E) and a significant reduction in OCT4 expression in EBs treated with the cocktail of factors (Figure 1D).
