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Fig. 4. Premature expansion and increased numbers of Ngn3⁺ pancreatic cells in Gdf11-deficient mice. Pancreatic Ngn3 expression (blue) detected by antisense RNA in-situ hybridization in Gdf11^+/+ (A,D,G), Gdf11^+/- (B,E,H) and Gdf11^-/- (C,F,I) embryos at the indicated embryonic stages. (G-I) Ngn3-expressing cells at E17.5 are marked by black arrowheads for clarity. (J,K) Pancreatic expression of insulin and glucagon (green) and Ngn3 (red) at E17.5 in mice with indicated genotypes. White arrowheads (K) mark three of the numerous Ngn3⁺ nuclei observed in Gdf11^-/- mice. (L) Quantification of Ngn3⁺ pancreatic nuclei per mm² tissue at E15.5 The increase of Ngn3⁺ pancreatic cells in Gdf1^+/– and Gdf11^-/- embryos compared with wild-type littermates is significant (asterisks) at P<0.05. Data from three to four embryos per genotype are presented as the average ± standard error of the mean. (M-O) Expression of HNF6 (black nuclei) in Gdf11^+/+, Gdf11^+/- and Gdf11^-/- mice at E17.5. Black arrowheads in N and O mark a subset of ductal cells surrounding lumen that are heavily stained by anti-HES1 antisera. (P) Quantification of HNF6⁺ nuclei per mm² tissue at E17.5. Data from three to four embryos per genotype are presented as the average ± standard error of the mean. Asterisks indicate that the difference in numbers of HNF6⁺ nuclei in Gdf11^+/- and Gdf11^-/- mice compared with Gdf11^+/+ mice is significant at P<0.05.

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