(A) Schematics of the Prox1 3′UTR and Prox1 3′UTR mutant reporters. The first nucleotide after the stop codon of mouse Prox1 mRNA is designated as number 1.
(B) HEK293T cells were transfected with one of the RNAs, one of the reporters, and the luciferase reporter system by using Lipofectamine 2000. RNAs included control RNA, miR-181a mimic and miR-181a inhibitor. Reporters included vector, Prox1 3′UTR, and Prox1 3′UTR mutant. The luciferase expression was determined as described under “Materials and Methods”. The results were normalized against internal control (R. reniformis luciferase) and further normalized against the results obtained from cultures transfected with control RNA in each group. MC: miR-181a mimic control; IC: miR-181a inhibitor control; MM: miR-181a mimic; IH: miR-181a inhibitor. *, p<0.05 compared to cells transfected with Prox1 3′UTR and control RNA, and treated with the same agonist. #, p<0.05 compared to cells transfected with Prox1 3′UTR and the same control RNA, and without agonist treatment. Data represent mean ± SEM of four independent experiments.