Immunostaining of Pancreas Samples

Pancreas samples were harvested from Hadh^lox/lox, SCHADKO, and β-SKO mice and immediately fixed in 4% paraformaldehyde solution overnight at 4 °C. Samples were then stored in PBS at 4 °C until processing and paraffin embedding. Immunostaining (5-μm tissue sections) for SCHAD and insulin was performed as previously described (14 (link)). For glucagon and SST staining, the sections were incubated overnight at 4 °C with mouse monoclonal anti-glucagon antibody (Sigma-Aldrich, G2654; 1:8000) or rat monoclonal anti-SST antibody (Abcam, ab30788; 1:500), followed by incubation for 1 h at room temperature with secondary antibody 1:400 from Jackson ImmunoResearch Laboratories (glucagon: #715-586-151; SST: #712-586-153).