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. 2010 Dec 14;5(12):e14311. doi: 10.1371/journal.pone.0014311

Figure 2. mHtt increases REST expression at the transcriptional level.

Figure 2

(A) Schematic representation of the human REST gene promoter. Exons a, b and c (open boxes) are downstream of the alternative promoters A, B and C, respectively. These exons are alternatively spliced to exon D (solid box) which contains the translation initiation site, with the adenosine of the ATG start codon being set as +1. A -3390-bp/+1 fragment was cloned upstream of the luciferase reporter gene in the pGL3-basic vector, to generate the 5′UTR-REST vector. (B) Activity of REST gene promoter in 6-day differentiated NG108 cells expressing Nter-15Q or Nter-142Q. 5′UTR-REST or pGL3-basic vectors were cotransfected with either Nter-15Q (open bar) or Nter-142Q (solid bar) and beta-galactosidase expression vectors into NG108 cells. Cells were harvested after 6 days of neuronal differentiation to measure the luciferase and beta-galactosidase activities. Luciferase activity was normalized to beta-galactosidase activity and expressed as fold increase above the control pGL3-basic. 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.