FIGURE 4.

Both of the nuclear translocation and catalytic activity of ChAT are required for CHT1 induction by ChAT. A, hChAT mRNA level in wt-hChAT-GFP (wt-hChAT), NLS2-(−)-hChAT-GFP (NLS2-(−)), inactive hChAT-GFP (Inactive), GFP alone, and nontransfected SH-SY5Y cells. Total RNA was extracted from the cultured cells and used as a template for a real time PCR assay. Values are the mean ± S.E. The non-parametric Mann-Whitney test was used for statistical analysis. ns represents no significant difference. B, real time RT-PCR analysis of CHT1 mRNA expression in wt-hChAT-GFP (wt-hChAT), NLS2-(−)-hChAT-GFP (NLS2-(−)), inactive hChAT-GFP (Inactive), GFP alone, and nontransfected SH-SY5Y cells. *, p < 0.05 by the non-parametric Mann-Whitney test. C, immunoblot of CHT1 protein expression. Nuclear (N) and cytoplasmic (C) fractions were obtained from each clone, and subjected to immunoblotting with anti-CHT1 and anti-β-actin antibodies. Immunoblotted β-actin was used as a loading control.