Ad.REST-VP16 activates neuronal differentiation marker genes in NT2 cells. Uninfected NT2 cells and NT2 cells infected with Ad, Ad.GFP or Ad.REST-VP16 were assayed for the expression of neuronal markers MyT1, NeuroD3, GluR, AchRB2, Scg10 and NaCh genes by RT–PCR (A). GAPDH was used as internal control gene. A lane with no mRNA was also used as a negative control. Uninfected NT2 cells and NT2 cells infected with Ad, Ad.GFP, Ad.REST or Ad.REST-VP16 were also analyzed by immunoblotting assays using anti-synapsin (B), anti-neurofilament 200 (C) and anti-class III β-tubulin antibodies (D). Anti-actin antibody was used as an internal control in these assays.