In situ ZnT8 186-194 MMr staining

from supplement: In-situ staining was performed as described (2). Briefly, unfixed, frozen sections were dried for 2 h, loaded with 1 µg of MMrs overnight at 4°C, washed gently with phosphate-buffered saline and fixed in 2% paraformaldehyde for 10 min. After a further wash, endogenous peroxidase activity was blocked with 0.3% H2O2. Sections were then incubated serially with rabbit anti-phycoerythrin, horseradish peroxidaseconjugated swine anti-rabbit and 3,3'-diaminobenzidine tetrahydrochloride substrate (Thermo Scientific). After a final wash, sections were counterstained with hematoxylin, dehydrated via sequential passages in 95-100% ethanol and xylene, mounted and analyzed using a Nikon Eclipse Ni microscope with NIS-Elements Analysis D software v4.40.)