DEV01411 Supplementary Material

Files in this Data Supplement:

• Supplemental Tables (PDF)
• Supplemental Figure 1 -

  Fig. S1. Reduced hnf6 expression in jagged 2/3 morphants. Quantitative RT-PCR using RNA from 48-hpf embryos shows that expression of hnf6, but not pdx1 is reduced 50% relative to tata binding protein (TBP) in jagged 2/3 morphants, when compared with mock-injected controls. P<0.01 (Student’s t-test) for hnf6 expression in jagged 2/3 morphants versus control.

• Supplemental Figure 2 -

  Fig. S2. Kidney defects in jagged 2/3 morphant larvae. (A-C) Histological cross-sections through the solitary glomerulus of 5-dpf wild-type (A) and jagged 2/3 morphant (B,C) larvae. Two types of defects were seen in the morphant glomerulus: a small glomerulus (B) was present in some larvae; in others, the glomerulus was disrupted by enlarged blood vessels (arrow) (C). (D-F) Histological cross-sections through the proximal tubules of 5-dpf wild-type (D) and jagged 2/3 morphant larvae. Precipitated material (arrowhead) is evident in the morphant tubules. Additionally, the tubular epithelium appears dysmorphic in comparison to wild type. Such defects were seen in all morphant larvae examined. Asterisk indicates proximal kidney tubules.

• Supplemental Figure 3 -

  Fig. S3. Ectopic duct formation in 72-hpf and 84-hpf Notch-activated larvae. (A-D) Confocal sections (6 µm) through the liver of wild-type (A,C) and Notch-activated (B,D) larvae processed for cytokeratin IHC. Larvae generated via heat-shock treatment as described in the text. Arrows indicate ectopic biliary ducts present in the 72-hpf (B) and the 84-hpf (D) Notch-activated larva that are also seen in 96-hpf larvae (Fig. 10I). 84-hpf specimens (C,D) were also stained with DAPI (blue) to reveal liver cell nuclei. (E,F) Confocal projections through the liver of wild-type (E) and Notch-activated (F) 120-hpf larvae processed for P-glycoprotein IHC. Note dysmorphic canaliculi in the Notch-activated larva.

• Supplemental Figure 4 -

  Fig. S4. Phylogenetic comparison of zebrafish jagged genes. This comparison was derived using a nearest-neighbor analysis of zebrafish and human ClustalW aligned protein sequences. This analysis shows that the zebrafish jagged 1 and jagged 3 genes are most closely related to human Jagged 1. Zebrafish jagged 3 shares the greatest similarity with human Jagged 1. Zebrafish jagged 2 is most closely related to human Jagged 2.