Figure 4: miR-142-5p and miR-130a-3p target the 3′UTRs of SOCS1 and PPARγ respectively.

(a) Heat map for gene-expression profiling of human M(IL-4) transduced with control ASO versus miR-142-5p ASO or control mimics versus miR-130-3p mimics. The mean fluorescence intensity was calculated as the average for three replicates, and the levels of expression were shown with log2-transformed values. The top 50 genes with the greatest changes compared with controls are shown (n=3). (b) Luciferase reporter assays for THP-1 cells transfected with pRL-TK vectors carrying SOCS1–3′ UTR versus SOCS1-mut-3′UTR or PPARγ-3′ UTR versus PPARγ-mut-3′ UTR in the absence or presence of the indicated miRNA mimics (mean±s.e.m., n=3 independent experiments; **P<0.01 compared to control by two-tailed Student’s t-test). (c,d) The human untreated and M(IL-4) were transduced with miR-142-5p mimics and miR-142-5p ASO, respectively. Expression of SOCS1 (c) mRNA and (d) protein was measured. The numbers above the blots present the intensity ratio of SOCS1/GAPDH analysed by ImageJ. (mean±s.e.m., n=3, *P<0.05; **P<0.01 by two-tailed Student’s t-test). (e,f) The human macrophages untreated and M(IL-4) were transduced with miR-130-3p ASO and miR-130-3p mimics, respectively. Expression of PPARγ (e) mRNA and (f) protein was measured (mean±s.e.m., n=3, *P<0.05; **P<0.01 by two-tailed Student’s t-test). (g) Time course of miR-142-5p expression in human macrophages as determined by qRT–PCR and SOCS1 protein level and STAT6 phosphorylation as determined by western blotting after IL-4 stimulation (mean±s.e.m., n=3). (h) Time course of miR-130a-3p expression in human macrophages as determined by qRT–PCR and PPARγ protein level as determined by western blotting after IL-4 stimulation (mean±s.e.m., n=3).