Figure 5. The IKKβ mRNA 3'-UTR is a direct target of miR-148a-3p in macrophages. (A) The IKKβ mRNA 3'-UTR sequence was aligned with the miR-148a-3p seed sequence. The sequence highlighted with red is potentially recognized. The mutated IKKβ mRNA 3'-UTR sequence is shown. (B) RAW264.7 cells were transfected with miR-148a-3p mimic or inhibitor together with a reporter containing wild type IKKβ (psiCHECK2-WT) or mutant IKKβ (psiCHECK2-MUT) as indicated. Data are shown as the relative ratio of firefly luciferase activity to Renilla luciferase activity. (C) EMSA for proteins extracted from Raw264.7 cells. (D) Co-immunoprecipitation with mouse monoclonal anti-Ago2 or preimmune IgG from extracts of Raw264.7 cells. mRNA levels in immunoprecipitates were determined by RT-qPCR. Data are shown as the mean±SD of at least three independent experiments. **p<0.01, ***p<0.001. ns indicates no significant difference.