(A) Western blot reveals a down-regulation of NRF-1 and NOS1 protein levels in NRF-1 shRNA-transfected neuron, whereas NOS2 protein levels were not affected. β-Actin served as a loading control. (B–C) N2a cells and primary neurons were transfected with shRNA against NRF-1 (light gray bars), or with empty vectors (black bars), or with scrambled shRNA (dark gray bars). NRF-2α served as a negative control. NRF-1, Nos1, COX2, and COX6c subunit mRNAs show significant decreases in shRNA-treated samples as compared to those with empty vectors, whereas Nos2, Nos3, and NRF-2α mRNA remained unchanged. N = 6 for each data point. *, P < 0.05; **, P < 0.01 as compared to empty vectors.