Fig. 4.

Csde1 interacts with ctnnb1 mRNA in ECs. (A) Pie chart depicting the distribution of Csde1 binding peaks. (B) RIP-seq reads distribution of ctnnb1 mRNA compared with control. Blue peaks indicate Csde1 binding sites. (C) RIP-qPCR analysis showing relative mRNA level of ctnnb1 in IgG and anti-Flag groups. Data are mean±s.d. **P<0.01 (two-tailed unpaired Student's t-test). ns, not significant. n=3 replicates. (D) Western blotting showing that endogenous Csde1 protein can be efficiently pulled down by ctnnb1 mRNA compared with anti-sense mRNA probe. (E) Western blotting showing the Flag-hCSDE1 protein from Flag-hCSDE1^WT- or Flag-hCSDE1^DN-transfected HEK293 cells pulled down with biotin-labeled ctnnb1 probe. (F) The endogenous ctnnb1 mRNA was detected by FISH, and overexpressed EGFP-Csde1 was detected by IF using an EGFP antibody in zebrafish at 36 hpf. The white dotted lines mark DA and CV regions. The bottom panels are magnifications of the dashed boxed areas (upper panels) showing the ECs with colocalization of EGFP-Csde1 and ctnnb1 mRNA (white squares). DA, dorsal aorta; CV, cardinal vein; NC, notochord. Scale bars: 15 μm.