Mitochondrial Profiling in Beta-Cell Subtypes

For comparing mitochondrial mass and activity, islets from 2–3 month-old MNYI mice (both males and females) were isolated and dissociated into single cells. They were then stained with MitoView^™ 633 or MitoView^™ 650 (Biotium) at 37 °C for 15 minutes in the presence of 20 mM glucose, followed by Flow-cytometry analysis (Ernst et al., 2023 (link)). These dyes assay the mitochondrial transmembrane potential and mitochondrial mass, respectively. The β-cells were gated into eYFP⁺ and eYFP⁻ β-cell subtypes in order to compare their MitoView^™ signals. For ADP/ATP ratio in purified β-cell subtypes, an EnzyLight^™ ADP/ATP ratio Assay Kit (BioAssay Syetems) was used, following the manufacturer’s protocols.