Figure 3.

Expression of DN:REST derepressed RE1/NRSE-regulated transcription in HD. a, Bdnf, synapsin-1, and M4 muscarinic cholinergic receptor mRNA levels were measured in native Hdh^7/7 and Hdh^109/109 cells after delivery of DN:REST or empty vector. Bdnf, synapsin-1, and M4 muscarinic cholinergic receptor mRNA levels were restored in Hdh^109/109 cells infected with DN:REST compared with untransfected cells and cells transfected with empty vector (− and E, respectively). mRNA levels of REST/NRSF and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), genes whose transcription is not regulated by the RE1/NRSE silencer, are not affected by DN:REST expression. Data are expressed as percentage of controls (100% was assigned to Hdh^7/7 cells) and represent the average of four independent experiments. *p < 0.05 versus Hdh^7/7 untreated cells (−); ^#p < 0.05 versus Hdh^109/109 untreated cells and cell with empty vector (− and E, respectively). b, Left, Bdnf exon II, III, and IV mRNA levels in Hdh^7/7 and Hdh^109/109 cells by semiquantitative radioactive RT-PCR. Right, Quantitative analyses of the BDNF mRNA levels. The peak densitometric areas relative to Bdnf II, III, and IV were normalized over the peak densitometric area of the β-actin band. Data are expressed as percentage of controls (100% was assigned to Hdh^7/7 untreated cells) and represent the average of three independent experiments. *p < 0.05 versus Hdh^7/7 cells; ^#p < 0.05 versus Hdh^109/109 untreated cells and cells with empty vector (− and E, respectively). c, Bdnf protein levels determined by ELISA on lysates from Hdh^7/7 and Hdh^109/109 cells and cells infected with DN:REST and empty vector. Data are the average of three independent experiments. *p < 0.05 versus Hdh^7/7 untreated cells (−). ^#p < 0.05 versus Hdh^109/109 untreated cells and cells with empty vector (− and E, respectively).