Insulin (Autostainer Link 48)

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Expression of PYY by cells within 5-μm sections of mouse and human pancreas was determined using a rabbit-derived antibody directed against PYY (1:50) and an anti-rabbit AlexaFluor® 488 secondary antibody (1:200). PYY localisation in islet cells was probed by co-staining with guinea pig anti-insulin (1:200), mouse anti-glucagon (1:100) and rat anti-somatostatin (1:50) antibodies and species-specific Alexa Fluor® 594 secondary antibodies (all at 1:200). Negative control sections were probed with AlexaFluor secondary antibodies in the absence of primary antibodies. Immunostained pancreas sections were visualised under a TE2000 fluorescent microscope (Nikon, Kanagawa, Japan) and analysed using Image J software (https://imagej.net/). The proportion of cells expressing PYY in each islet cell subpopulation was calculated by dividing the mean number of PYY-positive cells by the number of β-, α- or δ-cells per islet.

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