Figure 4

Loss of Nkx2.2 affects expression of known exocrine factors, Ela1, Spink3, and a novel secreted proteoglycan, Nepn. qRTPCR of (A) Ela1 mRNA was assessed at e12.5, e13.5, and e15.5, (F) Nepn mRNA was assessed at e13.5 and e15.5, and (M) Spink3 mRNA transcript levels were determined at e12.5 and e13.5, while all gene expression analyses compared wild type (grey bars) and Nkx2.2^-/- pancreata (black bars). In situ hybridization comparing wild type and Nkx2.2^-/- pancreas at e14.5 and e15.5 for Ela1 (B, C, D, and E), e12.5 and e14.5 for Nepn (G, H, J, and K), e12.5 and e15.5 for Spink3 (N, O, P, and Q). To show co-expression of Nepn in the exocrine compartment and confirm increased protein levels, Ela1 immunofluorescence was performed on adjacent sections (I and L;green). DAPI (C and E; blue) indicates nuclei. Error bars represent SEM and asterisks denote statistical significance < 0.05. Magnification 20×. Large dashed lines outline the pancreas at e12.5, e14.5, and e15.5. Small dashed lines outline areas of Nepn and Ela1 co-expression.