Fig. S2

Defective hepatocyte polarization in cloche mutant larvae. (A, B) Wild-type (A) and cloche^la1164 (clo) mutant (B) larvae visualized for Tg(flk1:EGFP)^s843 (green) and Alcam (red) expression at 60 hpf. The outlined areas are magnified and shown in A′ and B′. (C) Projected confocal image of a cloche^m378 (clo) mutant larva visualized for Tg(flk1:EGFP)^s843 (green), Alcam (red) and Prox1 (blue) expression at 80 hpf. There are almost no endothelial cells in the trunk of clo^m378 mutant larvae, but occasionally a few Tg(flk1:EGFP)^s843 positive cells are present (arrow). Alcam immunostaining is shown separately in C′ and is schematically presented in C″. It appears to exhibit extra small branches (arrowheads) compared to wild-type (Fig. 1B). (D) Number of small branches of the Alcam positive network in wild-type (average 22 branches per liver, n = 3, SD= 2.6) and cloche mutant (average 42 branches per liver, n= 4, SD = 5.7) livers at 80 hpf is shown in white and grey, respectively. (E) z-plane confocal images of the same clo^m378 mutant larva visualized for Prox1 (blue) and Alcam (red) expression. Alcam immunostaining is shown separately in E′. White arrowhead points to one of the small branches. (F) z-plane confocal images of a clo^m378 mutant larva visualized for Prox1 (blue) and Alcam (red) expression at 100 hpf. Alcam immunostaining is shown separately in F′.