Fig. 2. Pancreatic buds of PMycKO embryos. (A-Q) Wild-type and PMycKO pancreatic buds were analyzed by immunohistochemistry. (A,B) Anti-Myc staining of dorsal buds (black, arrow) at E12.5. (C,D) Immunofluorescence staining of dorsal buds with anti-PDX1 antibody (green) at E13.5. (E,F) Double-immunofluorescence staining of dorsal buds with anti-glucagon (red) and anti-PDX1 (green) at E10.5. (G) Quantification of glucagon⁺ cell area in dorsal buds of wild-type and PMycKO at E10.5. (H,I) Immunostaining of dorsal buds with anti-neurogenin 3 at E12.5 (black arrows). (K,L) Anti-PHH3 staining of pancreatic buds (black, arrows) at E12.5. (N,O) Immunostaining of pancreatic buds with anti-cyclin D1 antibody (black, arrows) at E12.5. (P,Q) Immunostaining of dorsal buds with anti-CDK4 antibody (cytoplasmic, brown, arrows) at E13.5. (J,M) Quantification of the number of neurogenin 3⁺- and PHH3⁺ cells in buds of E12.5 wild-type and PMycKO embryos are shown. Each histogram represents the mean±s.d. for ventral and dorsal buds of three embryos each. Insets in A, P and Q represent a 2x enlargement and a 4x enlargement in B, N and O. Background in C and D was stained by DAPI. Abbreviations: l, liver; st, stomach; vb, ventral bud; db, dorsal bud. Scale bar: 50 µm.