kdrl mutants developed similar hepatic injuries after acute ethanol treatment as the ZM-treated WT larvae. (A-D) Confocal three-dimensional projections showing the whole-liver intrahepatic vasculature in WT and kdrlum19 mutant larvae at 27 hpt. The intrahepatic vasculature is marked by Tg(kdrl:GFP) expression. Ventral view, anterior is to the top. Dashed line marks the liver. Scale bar: 40 μm. (E) Numbers (mean±s.e.m.) of intrahepatic vascular branches per liver in WT, kdrl heterozygotes and homozygous mutants at 27 hpt. (F,G) Numbers (mean±s.e.m.) of HSCs in WT, heterozygotes and mutants in different experimental groups at 0 hpt (F) and 27 hpt (G). (H,I) qPCR analyses showing the comparison of col1a1b, lamb1a, tgfb1a and acta2 expression in control and ethanol-treated WT and mutant livers at 0 and 27 hpt. Triplicates were performed. The results are represented as relative expression levels normalized to the housekeeping gene eef1a1l1 (mean±s.e.m.). (J,K) Percentages (mean±s.e.m.) of WT (black), kdrl heterozygotes (orange) and homozygous mutants (blue) with hepatic steatosis at 0 hpt (J) and 27 hpt (K) based on Oil Red O staining. Each experiment in E-G was repeated three times and the numbers of larvae analyzed are shown. Statistical significance in E,J,K was calculated by two-tailed Student's t-test, and in F-I by one-way ANOVA and Tukey's post-hoc test. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001; ns, not significant.
|
