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Fig. 1. NKX6.2 expression in the developing mouse pancreas. (A,F) lacZ expression, identified by X-gal staining in Nkx6.2^tlz/+ embryos. Immunofluorescence detection of NKX6.2 (B-E, G-J; confocal images) or ß-galactosidase (ß-gal) (K-M) together with various pancreatic markers. (N) Amylase only. (A) At E9.5, lacZ expression is detectable in the dorsal (dp) and ventral (vp) pancreatic anlagen. (B) Anti-ß-gal and anti-NKX6.2 staining show complete overlap in the stomach (s) and dorsal pancreas (dp) in Nkx6.2^tlz/+ embryos. In the pancreas, NKX6.2 is detected in a subset of PDX1⁺ cells (yellow cells in C), but not in glucagon⁺ (Glc) cells at E10.5 (arrowhead in D). NKX6.2 is also found in the stomach epithelium (C). At E12.5, NKX6.2 expression is largely restricted to the glucagon⁺ cells (E). At E15.5, lacZ expression is predominantly detected in the dorsal, but also in cells of the ventral pancreas (vp) (F), but also found in the distal stomach and duodenum (d) (F). (G,H) NKX6.2 is absent from both NGN3⁺ (G) and insulin⁺ (Ins) cells (arrowhead in H). By contrast, NKX6.2 colocalizes with glucagon and amylase (Amy) (I). Not all glucagon⁺ and amylase⁺ cells express NKX6.2 (arrowheads in I). At E15.5, the majority of NKX6.2⁺ cells co-express PDX1 (J). In contrast to NKX6.2, ß-gal occasionally colocalizes with NGN3 (open arrowhead in K) and insulin (open arrowhead in L) in Nkx6.2^tlz/+ embryos. However, the majority of NGN3⁺ or insulin⁺ cells are ß-gal negative (K, and arrowhead in L). Like NKX6.2, ß-gal is expressed in a subset of glucagon⁺ cells (open arrowhead indicates co-expressing cell; arrowhead indicates non-co-expressing cell in M) and in exocrine cells, as shown by co-localization of ß-gal and amylase on adjacent sections (arrow in M and N). Scale bar: 20 µm.

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