Fig 2: MIF is a direct target of miR-451. (A) Schematic of the MIF 3'UTR containing the miR-451 binding sites. (B) HUVECs were treated with hypoxia for the indicated time points. The relative expression level of MIF was detected by RT-qPCR. *P<0.05 and **P<0.01 vs. control group (0 h). (C) miR-451 level was measured in serum samples from cerebral ischemic patients (n=15) and normal participants (n=15) by RT-qPCR. **P<0.01, as indicated. (D) HUVECs were transfected with miR-451 mimics or mimics NC. Following 24 h, cells were treated with hypoxia for 6 h. The expression levels of MIF protein were determined by western blotting. **P<0.01 vs. mimics NC group. (E) HUVECs were transfected with miR-451 inhibitor or inhibitor NC. Following 24 h, cells were treated with normoxia for 6 h. The expression levels of MIF protein was determined by western blotting. **P<0.01 vs. inhibitor NC group. (F) Relative luciferase activity in HUVECs co-transfected with wild-type or mutant-type 3'UTR MIF reporter plasmids and miR-451 or miR-NC. All data are expressed as the mean ± standard deviation. **P<0.01, as indicated; ##P<0.01, as indicated. RT-qPCR, reverse transcription-quantitative polymerase chain reaction; MIF, migration inhibitory factor; HUVECs, human umbilical vein endothelial cells; miR, microRNA; NC, negative control; UTR, untranslated region; wt, wild-type; mut, mutant.