In situ hybridisation analyses of gc mRNA expression during hepatic organogenesis. gc expression is initiated weakly in the liver at 55 hpf (arrowhead, A), and levels of expression increase until around 72 hpf (B and C). Hepatic expression of gc is elevated from 72 hpf onwards (C and D). For comparison, visualisation of the digestive system at 60 hpf by foxA1 staining (E) and schematic representation (F). Amplification of gc from adult tissues reveals it is not only expressed in the liver, but additionally at lower levels also in the kidney and testes (G). (H) Time line depicting onset of gc expression with respect to timely expression of cp, fabp10 and gpia. Dorsal views, anterior to the top (A-F).

In situ hybridisation analyses of gc mRNA expression in hdac1 and cloche mutant embryos. Wild type embryos initiate gc expression from 55 hpf, and expression is maintained at 120 and 144 hpf throughout the liver (arrowhead; A and B). gc expression is initiated in the liver in a subset of hdac1 mutants at 120 hpf (arrowhead; C), and all mutant embryos at 144 hpf (arrowhead; D). (E–H) in situ hybridisation combining cp expression (red) and gc expression (blue). At 120 hpf a small subset of cp-positive hepatocytes express gc (arrowhead; F), while at 144 hpf gc is expressed throughout the cp-positive liver (H). Notably, a subset of hdac1 mutant embryos expressing gc in the liver also express gc in the retina (arrow; D). Cloche mutant embryos do not display defects in the initiation of gc expression in the liver at 60 hpf (arrowheads; I, K), or the heightening of gc expression at 72 hpf (arrowheads; J, L). (A–D, I–L) Dorsal views, anterior to the top; (E-H) Lateral views, anterior to the left.

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