fig3

Figure 3. Silencing of reporter constructs containing target 3’-UTRs for miR-100 and miR-125b. (A) Reporter constructs were generated in which the firefly luciferase open reading frame was fused to either plasmid 3’-UTR sequences (empty vector), 3’-UTR sequences containing three perfect binding sites for miR-100 or miR-125b (100/125 vector), or 3’-UTR sequences from candidate mRNA targets of miR-100 or miR-125b (experimental vector); (B) Quantification of relative luciferase levels after transfection of the indicated constructs into CC-CR cells or ΔmiR-100/miR-125b cells. Relative firefly luciferase levels were calculated by normalizing to a co-transfected internal control vector expressing Renilla luciferase. Data (mean ± SEM, n = 3) were analyzed using two-way ANOVA tests, with ^* indicating P < 0.05, ^** indicating P < 0.01, ^*** indicating P < 0.001, and ^**** indicating P < 0.0001. SEM: Scanning electron microscopy; ANOVA: analysis of variance.