miR‐467 mediates inhibition of TSP‐1 protein production by uncoupling mRNA from polysomes. A, Transfection of RF/6A with miR‐467 mimic results in inhibition of TSP‐1 production. Western blot, anti‐TSP‐1, and anti‐β‐actin antibodies. B, Inhibition of production of the reporter gene in cells cotransfected with miR‐467 mimic and the reporter construct expressing luciferase cDNA fused to TSP‐1 UTR cDNA. Luciferase activity in cell lysates, FI, fluorescence intensity; mean±SEM, n=4, *P<0.05. C and D, miR‐467 mimic induces uncoupling of TSP‐1 mRNA (C) and the chimeric luciferase‐TSP‐1 UTR mRNA (D) from polysomes and their association with the nonpolysomal fraction. mRNA of TSP‐1 and luciferase were detected by real‐time RT‐PCR in each RNA fraction. Mean±SEM, n=4, *P<0.05 compared with low‐glucose sample (=1), #P<0.05 compared with cells transfected with a control ribooligonucleotide (control r‐oligo). TSP indicates thrombospondin; UTR, untranslated region; SEM, standard error of mean; RT‐PCR, reverse‐transcription polymerase chain reaction.