FIG. 4.

In utero supplementation with Leu inhibits endocrine cell differentiation by downregulation of Ngn3 expression. A and B: Immunodetection and quantification of NGN3-positive cells (brown) in pancreatic sections that were prepared from E16 fetuses from Leu-supplemented pregnant dams and E16 fetuses from pregnant dams that were not supplemented with Leu. Scale bar = 12.5 μm. C: Real-time PCR quantification of Ngn3, Insm1, and amylase mRNA in fetal pancreata of E16 pups from Leu-supplemented pregnant dams and E16 pups from pregnant dams that were not supplemented with Leu. Data are mean ± SEM of at least three independent experiments. NS, not significant; **P < 0.01. (A high-quality digital representation of this figure is available in the online issue.)