Fig. 1.
SLIT2/3-encoded miR-218 regulates ROBO1 and ROBO2 expression. (A) Schematic of miR-218-1 and miR-218-2 encoding in intron 14 of human SLIT2 and SLIT3, respectively (adapted from the UCSC Genome Browser) (Kent et al., 2002). Plots show the conservation of regions of genomic DNA among 18 placental mammals calculated by phyloP (Siepel et al., 2005). Exons are indicated as black boxes. miR-218 is indicated by red boxes. The direction of transcription is indicated by arrows. (B) Conservation of the miR-218 binding site in ROBO1 and ROBO2 among various species. Sequences complementary to the miR-218 seed sequence are indicated in bold. (C) Luciferase assays in HeLa cells demonstrating regulation of human and zebrafish ROBO1 and ROBO2 by miR-218. miR-126 was utilized as a control. *, P<0.05 compared with UTRmut. (D) miR-218 overexpression leads to downregulation of ROBO1/2 mRNA in HUVEC. *, P<0.05 compared with control mimic. (E) Representative western blot demonstrates that ROBO1 levels are diminished in miR-218-transfected HUVEC. Data in C and D are mean + s.e.m.