A: Representative Western blot showing a band migrating at ∼90 kDa corresponding to neprilysin protein. A: Representative Western blot showing a band migrating at ∼90 kDa corresponding to neprilysin protein. Mouse islet proteins were extracted, subjected to SDS-PAGE, immunoblotted with a polyclonal rabbit anti-neprilysin antibody, and then visualized by enhanced chemiluminescence. Mouse kidney was used as a positive control (right lane). A smaller band, likely nonspecific, was present in kidney but not islets. Each lane represents islet lysate from one female mouse. NT, nontransgenic; T, hIAPP transgenic. B: Representative islets from a hIAPP transgenic mouse stained for neprilysin (green), insulin (red), and nuclei (blue). Neprilysin appears to be localized to both β-cells and non-β-cells, as seen in the merge, and is membranous/cytoplasmic, as seen in the insert. In a subpopulation of islet cells, neprilysin staining is more intense (arrows). An identical pattern was seen in nontransgenic islets. Sakeneh Zraika et al. Diabetes 2007;56:304-310 ©2007 by American Diabetes Association