The regulation of expression by hsa-miR-223 or hsa-miR-125b involves binding to specific sites in the 3′-UTRs of LMO2, PRDM1, or IRF4 transcripts. (A) Dual luciferase activity of reporter plasmids with the wild-type or mutated 3′-UTR of LMO2 fused to the luciferase gene upon hsa-miR-223 precursor cotransfection in HeLa cells. The same experiment performed for (B) PRDM1 or (C) IRF4 3′-UTR luciferase reporter plasmids after cotransfection with hsa-miR-125b precursor. ■ represents cotransfections with the corresponding miRNA precursor, and □, the cotransfection of the same reporter vector with the nontargeting control. Values are normalized to the value of each control, which is noted as 100%. Mutation of putative binding sites is expressed as MUT1 for the most 5′ site, MUT2 for the most 3′ site, and MUT1 + 2 for the combined mutation of both sites. Statistical comparisons by 2-tailed t test with Bonferroni correction between different constructs are represented as ↔. Statistical comparisons between the cotransfected miRNA and the nontargeting control for the same reporter vector are noted over the black bars. Significant differences with associated P values less than .05 are expressed as * and nonsignificant difference, as ns. Error bars correspond to the SEM.