FIGURE 5.

Neurofilament-L as a potential target promoter for activation by Pea3. (A) hNF-L promoter, which contains 2 major ets motifs, was cloned upstream of a luciferase reporter gene; (B) luciferase assays in SH-SY5Y human neuroblastoma cells transfected with or without mPea3, in the presence or absence of NGF stimulation; (C) luciferase assays in SH-SY5Y human neuroblastoma cells transfected with or without mPea3, in the presence or absence of retinoic acid (RA) stimulation; (D,E) SH-SY5Y cells are co-transfected with wild type NF-Lgene promoter driven luciferase reporter plasmid or with NF-L gene promoter which lacks putative Pea3 binding sites ets-1 (NFLΔ1-Luc) or ets-2 (NFLΔ2-Luc) or both (NFL Δ1Δ2-Luc). Relative luciferase activities were monitored in the presence (D) or absence (E) of 200ng of wild type Pea3. Luciferase activities represent reproducible three or four independent experiments, and data plotted for each condition were average of relative luciferase activity of triplicate samples.