Expression patterns of tie-1AS lncRNA. (A) The primer pair locations for quantitative PCR. Developmental expression profile of tie-1 (B) and tie-1AS (C) is shown. The total RNAs were extracted using Trizol followed by DNase I treatment. The RNAs were recovered by RNeasy kit (QIAGEN). (D) Tie-1 is expressed in the axial vessels () and blood islands (*). (E) Tie-1AS lncRNA is expressed in the axial vessels (), intermediate cell mass (*), otic vesicles (#), and brain (brace). (F) Transverse section of 24 hpf tie-1AS lncRNA embryo is shown. indicates the location of DA and PCV. (G) Luciferase activity in vivo (26 hpf) for a 3-kb fragment upstream of zebrafish tie-1AS lncRNA. The hRobo4 promoter is used as a positive control. **P < .01.