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In situ hybridization analysis shows a striking reciprocal pattern of REST expression to genes expressed within the developing nervous system, consistent with its role as a repressor (Schoenherr and Anderson, 1995; Chong et al., 1995).
We then examined the effect of mHtt on REST expression in NG108 cells expressing an amino-terminal fragment (Nter) corresponding to the first 171 amino acids of human Htt with 142Q (thereafter called Nter-142Q) or with 15Q as control (Nter-15Q).
Interestingly, however, we could detect REST protein in a small number of TuJ1+ neurons in the outmost MZ of the CP, corroborating β-gal staining, suggesting either that REST is re-expressed at later stages of differentiation or that REST expression has not yet been extinguished completely in these neurons.
Cytotoxicity and transfection efficiency at different siRNA doses were monitored by evaluating REST expression.
An inhibition of around 60% of REST expression was observed with both NP when associating 250 ng/mL of siRNA REST, as recommended for commercial reagents.
In Nter-15Q neuronal-like cells, Sp1/Sp3 double knockdown reduced REST expression at a level comparable to that of Sp3 knockdown alone (Fig. 6), thereby confirming that Sp3 is majorly involved in the basal REST expression in these cells.
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He is built as solidly as a trash compactor, and his resting expression is one of irritable disappointment.
Fiachra's resting expression was raked-in and solemn, like the heap of cinders left after a fire has gone out.
His resting expression already featuring eyes slightly widened as if by surprise, he seemed able to provide only one facial variation: a double up-flick of the eyebrows for emphasis, sometimes accompanied by a blink.
From the thick thatch of gray hair to the improbably long and thrusting chin, Kerry's visage is immense and, in its implacable resting expression, resembles one of the monolithic heads that rise from the loam of Easter Island.
Kellner compares him to a "domesticated werewolf," placid but intense, with a resting expression that was an unnerving "blank stare," and "disfigured by two very big and very ugly protruding front teeth, like squares of broken crockery".
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