qRT-PCR analysis of developmental REST expression. (A) Electrophoresis gel of RT-PCR products showing relative levels of REST (top) and β-actin (bottom) expression at 12 hours post-fertilization and then daily from 1 to 7 dpf. (B) Graph showing normalized REST expression levels relative to β-actin controls as assayed by qRT-PCR. This analysis shows that REST expression increases significantly at 6 dpf, a time point correlated with reductions in skeletal muscle expression in NRSE-containing enhancer trap lines. Relative expression analyses were performed using 2-ΔΔCT methods; the experiment was repeated a total of three times, with averages and standard deviations shown per time point.