|
Fig. 2 Homozygous knockout mutants are sensitive to DEB treatment.
(A) Embryos obtained from inbreeding heterozygous knockouts of fancd1 (hg45; 0.9 μg/mL DEB), fancd2 (hg47; 0.9 μg/mL DEB), fanci (hg54; 0.65 μg/mL DEB), fancj (hg56 and hg57; 0.6 μg/mL DEB), fancn (hg62; 0.8 μg/mL DEB), fancp (hg66; 0.9 μg/mL DEB) and fanct (hg70; 0.8 μg/mL DEB) were treated at indicated DEB concentrations between 4–72 hpf. Treated embryos were classified based on severity of morphological changes observed into three phenotypic groups: normal (WT appearance), moderate (slight body curvature and minor edema) and severe (emaciated appearance, severe body curvature and large edema). An example image of DEB treated embryo (72 hpf) for each group is shown on the left. Distribution of each phenotypic group for a given genotype are displayed as stacked bar chart. The segments in bar show percent of embryos in each morphological group: normal (white), moderate (light blue), severe (dark blue). The number in each segment depicts the number of embryos for a given phenotypic group. (B) Maternal WT fanca transcript rescues embryos from DEB hypersensitivity. Embryos obtained from indicated fanca_hg41 breeding were treated with DEB (0.8 μg/mL). Representative images show untreated and DEB treated embryos. (C) Embryos generated from inbreeding of homozygous knockouts of fancb (hg42; 0.8 μg/mL DEB), fanco (hg65; 0.8 μg/mL DEB) and fancq (hg69; 0.5 μg/mL DEB) were treated at indicated DEB concentrations. Representative images show untreated and DEB treated embryos.