Total RNAs were prepared by using RNeasy mini-columns (Qiagen, Valencia, CA, USA) according to the manufacturer’s protocols. cDNAs were synthesized by SuperScript III first-strand synthesis kit (Life Technologies). The pre-validated primer sets for neural markers were purchased from Real Time Primers, LLC (Elkins Park, PA, USA). The amplification was done in a final volume of 20 μl under the following conditions: 15 min at 95°C and then 55 cycles at 95°C for 10 seconds, 58°C for 45 seconds, and 72°C for 45 seconds. The sizes of qPCR products were confirmed by agarose gel electrophoresis. Biorad iCycler was used to determine Ct values for each sample. Gene expression levels were normalized against β-actin levels in each sample and the fold changes were calculated by setting the expression levels of each genes in undifferentiated control ReN-G cells as 1. The following are the neuronal and glial marker gene names and PCR product sizes: NCAM1 (neural cell adhesion molecule 1, 174 bp), SYT5 (synaptotagmin V, 171 bp), SLC17A7 (VGLUT1, solute carrier family 17 (sodium-dependent inorganic phosphate cotransporter) member 7, 162 bp), GRIN2A (glutamate
