Fig. S2

Fig. S2 PGE2 affects Pancreas Development (Related to Figure 2) (A) In situ hybridization for carboxypeptidase A (cpa), an exocrine pancreas-specific marker, reveals that embryonic pancreas size is regulated by PGE2. Exposure to dmPGE2 from 48-96 hpf leads to a dramatic increase in embryonic pancreas size at 96 hpf, whereas exposure to Indomethacin severely abrogates embryonic pancreas development. Dorsal view. (B) Histological analysis reveals that modulation of PGE2 levels does not alter liver morphology (n = 5 / treatment). TUNEL analysis shows no apoptosis following modulation of prostaglandin activity (n = 5 / treatment). (C) Representative view of ck19 immunohistochemistry of the developing pancreas at 96 hpf in all treatment conditions. Low levels of ck19 are broadly expressed, and higher expression is just beginning to appear in developing pancreas ductules. (D) Pharmacologic blockade of PGE2 synthetic enzymes interferes with exocrine pancreas development, as visualized by in situ hybridization for trypsin. Blockade of cox1a with SC-560 or cox2a with NS-398 results in a smaller pancreas. In contrast, blockade of the PGE2 degrading enzyme pdgh with Cay10397 results in a larger embryonic pancreas. (n > 50 / treatment). Dorsal view. (E) Morpholino-mediated knockdown of PGE2 synthetic enzyme pges abrogates pancreas development, as visualized by in situ hybridization for trypsin expression. (n > 30 per condition). Left lateral view. (F) In situ hybridization for intestinal fatty acid binding protein (ifabp), a marker specific for the developing gut endoderm, reveals that PGE2 modulates the size of the embryonic intestine. Exposure to dmPGE2 from 48-96 hpf enlarges the developing intestine, whereas exposure to Indomethacin diminishes the size of the embryonic intestine. (n > 30 / treatment). Dorsal view. (G) Quantification of GFP+ cells by FACS of gut:GFP embryos at 96 hpf corroborates the findings of in situ hybridization in (F). Exposure to dmPGE2 from 48-96 hpf enlarges the developing intestine, whereas exposure to Indo abrogates intestinal development. (Data are represented as mean ± SEM).