Fig. 3

Zebrafish phenotype associated with INF2 expression and Dia activity. Control MO and INF2 ATG MO and in vitro transcribed hINF2 mRNAs were co-injected into zebrafish embryos at the single-cell stage. a, b, c. 400 zebrafish expressing different forms of INF2 (50 per group) were categorized and quantified based on different gross phenotype and histology at 96 h post fertilization (hpf). Phenotype 0: grossly and histologically identical with wt fish at 96 hpf; phenotype 1: significant pericardia edema, slightly disorganized sarcomeres at the tip of the tails, without significant tail curling or dorsalization. Histologically collapsing or underdeveloped glomerular tufts.; phenotype 2: Diffuse edema with prominent yolk, significant bending and dorsalization. Histologically, underdeveloped glomerulus without open capillary tuft.; phenotype 3: dorsalized embryo with shortened tail, diffuse edema, underdeveloped pronephros, and distended tubules. Statistical significance, χ² test: *, p < 0.05 vs. C-MO only; ^, p < 0.05 vs. ATG-MO only. d. Expression of hINF2 protein (Flag-tagged) was confirmed by western blot in 50 zebrafish in each group. e. Rho activity in zebrafish with expression of different forms of INF2 was measured by Rhotekin RBD pulldown assay. f. Dia activity in zebrafish with different manipulations of INF2 was measured and quantified as the ratio of membranous associated Dia2 (Membrane/Total Dia2).