Figure 2

A pool of miRNAs dysregulated in sALS regulates the activity of a reporter linked to NFL 3′UTR. Reporter gene assay was performed transfecting HEK293T cells with pre-miRNAs and a reporter construct expressing firefly luciferase coupled to NFL 3′UTR-S (white bar), -M (grey bar) or -L (black bar). The results of firefly luciferase activity were normalized with Renilla reporter luciferase activity and with the effect of each miRNA on the luciferase mRNA alone (pmirGLO vector control). Data are expressed as relative change and show positive values as up-regulation and negative values as down-regulation. Experiments were performed in triplicate. Result are shown as mean ± SEM (t-test: *** = p < 0.001; ** = p < 0.01, relative to the pmirGLO vector control). Note that not all miRNA have predicted interaction sites in all lengths of NFL 3′UTR, only data for which a predicted site is present are shown. In addition, only miRNAs with correlation among the results of the reporter gene assay and the expression observed in sALS are presented.