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. 2008 Dec 31;28(53):14341–14346. doi: 10.1523/JNEUROSCI.2390-08.2008

Figure 1.

Figure 1.

Several REST-regulated miRNA levels change with HD progression, and miR-9/9* targets a silencing complex. A, Longitudinal expression profiles of REST-regulated miRNAs in HD and healthy control cortex (BA4) HD grade 1, 2, and 3/4 as measured by QPCR. Mature miRNA levels in HD brain relative to controls (*p < 0.05, **p < 0.01). B, C, miR-9/9* reduces expression of reporter proteins containing the 3′ UTR of REST or CoREST. B, HEK 293 cells were cotransfected with a vector expressing miR-9/9* (mU6 miR-9/9*) and REST 3′ UTR luciferase reporters. Relative luciferase activity was measured 24 h later. Increasing doses of mU6 miR-9/9* relative to target caused a dose-dependent decrease in relative luciferase. Control vectors containing a single site with perfect complementarity to mature miR-9 (miR-9-PT) or miR-9* (miR-9*-PT) were included as positive controls (n = 3). C, Experiments as in (B) with CoREST 3′ UTR luciferase reporters (n = 3). D, E, Transfection of HEK 293 cells with mU6 miR-9/9* reduces both REST and CoREST protein expression levels 24 h later. D, Representative Western blots and densitometry for (E) REST (n = 5) and CoREST (n = 5). Histograms show mean ±SEM after normalization to β-actin levels. *p < 0.05, **p < 0.01.