Loss-of-function of Leg1 blocks the liver bud formation.
(A) Loss of maternal zygotic Leg1a affects the liver development. Embryos were treated with or without UV25 at 24hpf, WISH was performed to assess the development of the liver (fabp10a), exocrine (trypsin) and endocrine (insulin) pancreas and intestine (fabp2a) in the maternal-zygotic leg1azju1(mu) embryos at 3.5 dpf. At least three independent WISH was performed, each time with 24–31 embryos for each sample, and representative embryos were shown. (B) WISH using prox1 and hhex to examine liver bud formation at 30 hpf in embryos treated with or without UV25 treatment at 24 hpf. Representative pictures in each group are presented. The number of embryos exhibiting the phenotype over total embryos examined are shown in the bracket. Red arrow: liver bud, blue arrow: pancreatic bud. (C and D) Images (C) and statistical analysis (D) of PH3 immunostaining to compare cell proliferation of hepatocytes in WT and maternal-zygotic leg1azju1 embryos (mu) after UV25 treatment. BHMT is an enzyme highly expressed in the liver and was used to mark out the hepatocytes. DAPI was used to stain the nuclei. Red arrows indicate PH3 positive cells in the liver. PH3 positive cells in the neural tube in WT and the mutant were also recorded in parallel. *, p<0.05. in, intestinal tube, lv, liver, nt, neural tube.