Figure 4.

miR-133b targets FOXC1 in OS. (A) Targetscan software predicted that FOXC1 was a putative target gene of miR-133b, and that (B) their targeting relationship was evolutionally conserved. (C and D) WT- or MT-FOXC1 3′UTR was cloned into a pMIR-GLOTM Luciferase vector downstream of the F. luciferase coding region. (E) Luciferase activity was significantly decreased in U2OS cells co-transfected with miR-133b mimic and WT-FOXC1-3′UTR plasmid but not in cells co-transfected with miR-133b mimic and MT-FOXC1-3′UTR plasmid. **P<0.01 vs. miR-NC. FOXC1, forkhead box C1; miR, microRNA; OS, osteosarcoma; WT, wild type; MT, mutant type; 3′UTR, 3′ untranslated region; NC, negative control; SV40, simian vacuolating virus 40; R. luciferase, Renilla-glo luciferase; F. luciferase, firefly luciferase; HSV TK, herpes simplex virus thymidine kinase promoter; Sgf1, XhoI, PmeI and NotI, restriction sites.