FIG. 2.

Molecular profiling of the temporal expression of expected genes in the 5-stage of mouse ES cell differentiation toward mDAergic neurons. (A) Relative mRNA expression levels for pluripotent markers, Nanog and Oct4, neural progenitor markers, Pax6, Sox1, Nestin, and Ncad, ventral midbrain (VM) markers, En1, Otx2, Foxa2, and Lmx1a as well as pan-neuronal markers, Tau and Dcx. Mean±SEM; n=6. All markers were compared to the stage, where the expression was first detected by real-time PCR. Gene expression was considered as not detected (ND) when ΔCt >14 cycles. (B) Relative mRNA levels and percentage of Th+neurons in stage V show 2 different experimental outcomes: one positive (+), where Th+ neurons were found, and another negative (−), where very few Th+ neurons were counted (see also Supplementary Table S2). Mean±SEM; n=3. **P<0.01 significantly different from (−) experimental outcome by Student t-test. Th, tyrosine hydroxylase; SEM, standard error of the mean; PCR, polymerase chain reaction.