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The sections were first incubated overnight in peroxidase-conjugated sheep anti-digoxigenin antibody Fab fragments, and then the Glp1r hybridization signal was amplified using the TSA Plus DIG Kit (Akoya Biosciences, Cat# NEL748E001KT) for 30 min, applying the DIG amplification reagent at 1:500 dilution in 0.05 M Tris (pH 7.6) containing 0.01% H 2 O 2