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Fig. 1 Deletion of sav1 causes larval lethality and abnormal gallbladder development. (A) Wild-type and mutant alleles of zebrafish sav1. sav1 mutants contain an 11 bp insertion (red), causing a frameshift and early stop codon (red box). (B) Morphology of embryos at 8 dpf. Dashed red lines on higher magnification images outline the gallbladder. Pan, pancreas; Gb, gallbladder. (C) Quantification of gallbladder area shown in B; gallbladder location is indicated by red circles. Unpaired t-test with equal s.d., P<0.0001. n=38, sav1+/+ or +/−; n=12, sav1−/−. Data are mean±s.d. (D) BODIPY FL C5 gallbladder uptake and ratiometric analysis at 8 dpf. Sample number (n) indicated within the graph. (E,E′) Hematoxylin and Eosin stained paraffin sections of normal gallbladder and liver in wild-type and sav1−/− mutants. Gb, gallbladder. Scale bars: 100 μm. (F,F′) Adjacent sections stained for annexin A4 confirm abnormal biliary morphology. Gb, gallbladder; IBD, intrahepatic bile duct. n=5 for both wild-type and sav1−/− paraffin histology.