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Fig. 3. Quantitative transcription profile of hMSC, hmNSCs and terminal differentiated hmNSCs into glial and neuronal cell types. (A) Representative real-time RT-PCR analysis using the LightCycler® technique. Plot of the fluorescence versus the cycle number obtained from SYBR Green detection of serially diluted FN1 mRNA (encoding fibronectin) (left). The horizontal line represents the position of the threshold. The standard curve obtained by plotting cycle number of crossing points versus dilution factor is shown (center), in addition to melting curve analysis showing the specificity of the amplified PCR product (right). (B) Quantitative real-time RT-PCR analyses of mesodermal genes (FN1), proneural genes (SOX1, OTX1, NeuroD1, Neurog2, MSl1), NSC marker genes (NES), glial genes (GFAP, MBP) and neuronal genes (TUBB4/III, SNCA, NTRK1, TH) as well as OCT-4 as a marker for pluripotency in hMSC, hmNSCs and differentiated hmNSCs, respectively, by the neuronal induction protocol for 14 days. Expression levels are expressed relative to the housekeeping gene HMBS (hydroxymethylbilane synthase). For primers, complete names of genes and melting curve analyses demonstrating the specificity of amplified PCR products see Table 1. #, P<0.05; ##, P<0.01 when compared to mRNA levels in hMSCs; +, P<0.05; ++, P<0.01 when compared to mRNA levels in hmNSCs.

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