In vitro functional analysis of APP A. Schematic representation (not to scale) of the luciferase reporter construct used in this study. TK; thymidine kinase promoter, AAAA(n); PolyA site B. HEK293T cells were transfected with 50nM pre-miRs (as indicated) as well as a reporter construct containing the 3'UTR of hAPP. The cells were lysed 24h post-transfection and luciferase signal was measured. Signals were normalization for transfection efficiency and graph represents the luciferase signals compared to the scrambled control (SCR). Statistical significance was assessed by Student paired t-test. (* = p < 0.05, ** = p < 0.01, *** = p < 0.001) C. and D. Neuro2A cells (C) and HeLa cells (D) were transfected with 50nM pre-miRNAs (as indicated). The cells were lysed 48h post-transfection and western blotting was performed. Representative (n = 3, in triplicate) western blots are shown. The ratios of the APP/β-Actin signals are presented. Measurements were normalized to the scrambled control (SCR). Statistical significance was assessed by Student's paired t-test. (* = p < 0.05, ** = p < 0.01, *** = p < 0.001).