Fig. 2: Analysis of cellular oncomiR-targeting site bearing miRDRELs.

a Two consecutive cellular miRNA-targeting sites of let-7, miR-21, and miR-122 in ubiquitin-conjugating enzyme E2 R1 (CDC34), protein phosphatase 1 regulatory subunit 3B (PPP1R3B), SATB homeobox 1 (SATB1), and glycogen synthase 1 (GYS1) mRNAs were inserted into the firefly 3′UTR of luc-miRDREL. The relative distance and location of each miRNA-targeting site within the 3′UTR are depicted in panel (b). NT, natural miRNA targets. c–e Luciferase activities of each luc-miRDREL were compared to that of the target site null control. Repression of luciferase activity indicated that each luc-miRDREL was targeted by endogenously expressed let-7a, miR-21, or miR-122 in various carcinomas. The data represent the mean values of four independent experiments (n = 4). Error bars in the graph represent ±standard deviation, and the P-values are based on a comparison of the control to luc-NT-miRDRELs. **P < 0.01, ***P < 0.001. f, g Analysis of the cellular miR-122-targeting site bearing luc-GYS1-miRDREL in cells not expressing miR-122. The luciferase activity of luc-GYS1-miRDREL was not changed in the NCI-H1299 or PANC-1 cells, which do not express miR-122. The luciferase activity of luc-GYS1-miRDREL was compared to that of the miRNA-targeting site null control. The data represent the mean values of three independent experiments (n = 3). Error bars in the graph represent ±standard deviation. NS, not significant.