Figure 4

In-vitro analysis of HD-associated miR-128a gene targets. The regulation of the 3’-UTR activity by miR-128 for (A) HIP-1, (B) HTT, (C) SP-1, and (D) GRM-5 was evaluated in 293-FT cells. 293-FT cells were co-transfected with 10 nM pre-miR-128a or negative control (NC) as well as luciferase reporter constructs containing the WT or mutant (MUT) 3’- predicted miR-128a binding site for each of the genes. Relative luciferase activity data were normalized as a percentage of plasmid-only control levels (100%) for each 3’-UTR construct. One-way ANOVA followed by Tukey’s post-hoc testing confirmed that miR-128a exposure resulted in significant reduction (p < 0.001) in 3’-UTR activity for each WT construct (as compared to NC); no significant differences between treatments were detected in MUT constructs.