Fig. 5.

Relative expression of DDX3X and CCNE1 in Ishikawa cells after overexpression of miR-181a (pre-miR-181a), or PreNC determined by quantitative RT-PCR (A and D) and Western blot analysis (B). C, Firefly luciferase assay with pZEX-MT01 construct carrying a 3′-UTR fragments of DDX3X in Ishikawa cells cotransfected with firefly luciferase reporter, pre-miR-181a, or PreNC. The ratio of firefly to renilla was determined and reported as relative luciferase activity compared with empty vector with levels in PreNC independently set at 1. The results present mean ± sem of three sets of independent experiments performed in duplicate and analyzed using nonparametric Student's t test with P values presented as indicated by corresponding line. Sequence alignment of miR-181a seed region and DDX3X mRNA target site at 3′-UTR with the coordinated positions is shown at the top of each graph. E, Relative expression of BCL2 in Ishikawa cells after overexpression of miR-181a (Pre-miR-181a) as compared to its control (PreNC) and depletion of miR-181a (Anti-miR-181a) as compared to its control (AntiNC) by quantitative RT-PCR.