MiRNAs hsa-miR-374a-5p, hsa-miR-542-3p, hsa-miR-216a-5p, hsa-miR-19b-3p, and hsa-miR-340-5p target the MID1 3’-UTR.

<p>(A) HEK293T cells were transfected with a pool of miRNA mimics (hsa-miR-374a-5p, hsa-miR-542-3p, hsa-miR-216a-5p, hsa-miR-19b-3p, and hsa-miR-340-5p), a pool of miRNA inhibitors (hsa-miR-374a-5p, hsa-miR-542-3p, hsa-miR-216a-5p, hsa-miR-19b-3p, and hsa-miR-340-5p), or a non-specific control siRNA. 24 hours later the same cells were transfected with a reporter construct carrying renilla luciferase fused to the MID1 3’-UTR sequence (bp 1–1352) as well as firefly luciferase, which is used for normalization. Relative light units of renilla normalized to firefly luciferase are shown. Columns represent mean values +/- SEM (* p < 0.001, n = 13). (B-F) Cotransfection of HEK293T cells with miRNA mimics in combination with reporter constructs carrying renilla luciferase fused to the MID1 3’-UTR sequence (bp 1–1352), in which the seed regions of the specific miRNAs have been mutated (MUT). As in (A) firefly luciferase was used for normalization. The resulting data were normalized to the mean value for a control using the non-mutant (WT) MID1 reporter construct described in (A). Columns represent mean values +/- SEM (* p < 0.01, n = 18).