MicroRNA-98 and let-7 target CIS 3′-UTR causing translational suppression. A, The schematic of CIS mRNA shows two potential binding sites in the 3′-UTR for miR-98 and let-7. B, Binding of miR-98 and let-7 to the two potential binding sites in the CIS 3′-UTR results in translational suppression. CIS 3′-UTR sequence covering both potential binding sites was inserted into the pMIR-REPORT luciferase plasmid. A control plasmid with the mutant 3′-UTR sequence was also generated. H69 cells were transfected with the constructs and treated with the anti-miRs or miRNA precursors followed by luciferase analysis. C, miR-98 and let-7-mediated translational suppression of CIS requires both the potential binding sites for miR-98 and let-7 in the 3′-UTR. The two binding sites for miR-98 and/or let-7 in the CIS 3′-UTR were inserted separately to the pMIR-REPORT luciferase plasmid (3′-UTR Site A vs 3′-UTR Site B). Control plasmids with the mutant 3′-UTR sequence in each potential binding site were also generated. H69 cells were transfected with the reporter constructs for 24 h followed by luciferase analysis. Mut = mutant; *, p < 0.05 ANOVA vs the controls; #, p < 0.05 ANOVA vs CIS 3′-UTR transfected cells.