Fig. 5

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 kdrl^um19 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??m. (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.