Effects of zNgBR and zNogo-B MO-injection on ISV formation. (A) zNgBR MO blocks ISV formation. Whole-mount fli1a- in situ staining (blue; Fli1a-WISH) was performed to localize the ISV at 24 hpf embryos. Top panels are the images of whole embryos at 24 hpf, and bottom panels are magnified images of the trunk region, showing WT (uninjected), Ctrl MO- (8 ng control MO), MO1- (8 ng ATG MO), and MO3- (8 ng splice MO) injected embryos. Asterisk indicates location of ISV. (B) Quantification of embryos with ISV defects (missing more than 5 ISV). n, embryo number in each group. (C) zNogo-B MO causes ISV formation defects. Fli1a-WISH (blue) was performed to localize the ISV at 24 hpf embryos. Top panels are the images of whole embryos at 24 hpf, and bottom panels are magnified images of the trunk region, showing WT (uninjected), MO1- (8 ng ATG MO), and MO2- (8 ng splice MO) injected embryos. Asterisk indicates location of ISV. (D) Quantification of embryos with ISV defects. n, embryo number in each group. Severe defect, missing more than 3 ISV in trunk region; mild defect, ISV orientation defect. Anterior is to the left.