Quantitative PCR analysis using a microarray was performed as described in our previous publication (Tyler & Allan, 2013). Briefly, total RNA was extracted from cell pellets derived from plates from either proliferating NPCs or differentiated NPCs using the RNeasy Mini Kit and a QIAshredder homogenizer (Qiagen; 74134, 79654). Each plate contained approximately 1.2 × 106 cells. The mRNA concentration was determined using a NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies); all RNA used for PCR analysis had a 260/280 absorbance ratio of ~2.0. Approximately 750 ng of isolated mRNA was converted to cDNA using the RT2 First Strand Kit (SABiosciences; 330522). PCR analysis was performed according to the manufacturer's instructions on an ABI 7300 Real-Time PCR System (Applied Biosystems) using the RT2 Profiler Mouse Neural Stem Cell and Neurogenesis PCR Array (SABiosciences; PAMM-404). Cycle threshold (CT) values for each gene were normalized to an average CT value from two housekeeping genes (Gusb and Hprt). Although Gapdh, β-actin, and Hsp90ab were also on the array and available for use as housekeeping genes, our lab has found that prenatal alcohol exposure impacts the expression
