Figure 1

Search for miRNAs targeting Prox1. (A) Seed matches between the 3' UTR of Prox1 and miR-466, miR-4305, and miR-4795-5p. The coordinates for miR-466 target sites 1 and 2, as well as the target site for miR-4305 and miR-4795-5p, are shown in parentheses (GenBank accession number NM_002763). (B) Cross-species sequence alignment of the 3' UTR of Prox1. The shaded boxes are putative miR-466, miR-4305, and miR-4795-5p target sites. Con indicates conserved residues. Alignment of the Prox1 3' UTR shows the sequence conservation among the human (has), chimpanzee (ptr), mouse (mmu), rat (rno), rabbit (ocu), and dog (cfa). Numbers indicate the nucleotide position after the stop codon of Prox1. (C) Direct targeting of the Prox1 3' UTR by miR-466, miR-4305, and miR-4795-5p. Luciferase activity was measured in HEK293T cells co-transfected with psiC-Prox1 and the miR-466, miR-4305, or miR-4795-5p mimics. miR-181a, which was reported to target Prox1, was used as a positive control. Luciferase activity was normalized using internal firefly luciferase activity, and expressed as a ratio to the luciferase activity obtained from the scrambled control-transfected cells. Error bars indicate SDs (n = 3 per experiment). *P < 0.05. †P < 0.01.