Figure 4. NF-Kappa B is not directly involved in the activation of REST promoter by mHtt.

On the left, schematic representations of the three REST promoters A, B and C, with or without mutation(s) in NF-Kappa B binding sites. Numbers indicate the relative positions with respect to the ATG start codon. The black lines represent the REST promoter regions, the open boxes the three exons (a, b, c) and the dotted lines the deletion between promoter regions and the luciferase gene (luc.) (light grey boxes). Dark grey boxes refer to the location of NF-Kappa B binding sites, while X indicates that these sites were mutated. On the right, luciferase activities of mutated promoters. Each mutated construct was cotransfected with Nter-15Q (open bar) or Nter-142Q (solid bar) and beta-galactosidase expression vectors into NG108 cells. After 6 days of neuronal differentiation, cells were harvested to measure the luciferase and beta-galactosidase activities. Luciferase activity was normalized to beta-galactosidase activity and expressed as fold increase above the activity of construct A, co-expressed with Nter-15Q. Data are the mean values ± sem of 3 independent experiments performed in triplicate. ***P<0,001, Nter-15Q-expressing cells vs. Nter-142Q-expressing cells.