Fig. 4

Yap but not Taz drives sav1?/? phenotypes. (A-A?) Whole-mount GFP staining of ctgfa:d2GFP wild-type and sav1?/? embryos. Gb, gallbladder. Numbers indicate the ratio of fish that displayed the phenotype/total fish analyzed. Scale bar: 100 ?m. (B,B?) Images of ctgfa:d2GFP+ wild-type and sav1?/? larvae; dashed yellow line outlines the liver. Scale bar: 100 ?m. (C) Mean pixel intensity of ctgfa:d2GFP reporter in livers as outlined in B: n=16 wild-type; n=9 sav1?/?. Data are mean±s.d. Unpaired t-test with equal s.d. resulted in P<0.0001. (D-E?) Immunofluorescent staining of paraffin wax-embedded sections for Yap and Taz in sav1?/? and wild-type siblings, and colocalization with the biliary marker annexin A4. Gb, gallbladder; IBD, intrahepatic bile duct. Scale bar: 100 ?m. (F-F?) Whole-mount staining for annexin A4, EdU and Taz in wild-type larvae. Gb, gallbladder; IBD, intrahepatic bile duct. (G-G?) Whole-mount analysis of sav1?/? larvae using the markers described in F (n=6). (H-H?) Whole-mount analysis of sav1?/?;yap?/? larvae using the markers described in F (n=8). Scale bar: 100 ?m. (I-K?) Repeated whole-mount analysis for the markers as described in F-H (although n=5 for sav1?/?;taz?/? embryos). Scale bar: 100 ?m. (L) Toluidine Blue stained sections of liver and gallbladder in wild-type, sav1?/? and sav1?/?;yap?/? larvae at 10?dpf (n=9). Gb, gallbladder; CBD, common bile duct. n=7 wild type; n=5 sav1?/?; n=4 sav1?/?;yap?/?. Scale bars: 100 ?m. (M) Ratiometric analysis at 8?dpf of BODIPY FL C5 gallbladder uptake in larvae from a sav1+/?;yap+/? incross. Analyses performed at 8?dpf unless otherwise indicated.