Quantifying Mitochondrial Function in Beige Adipocytes

Cells were seeded and differentiated on XF96 assay plates (Seahorse Biosciences, North Billerica, MA, United States) to white, active beige, or inactive beige using the protocols as described in Section 2.3. The OCR and ECAR were measured with XF96 oximeter (Seahorse Biosciences). Dibutyryl-cAMP stimulated OCR and ECAR, etomoxir-resistant (ETO-R) OCR, and stimulated proton leak OCR were quantified using previously utilized protocols (Kristóf et al., 2015 (link); Arianti et al., 2021 (link); Nagy et al., 2022 (link)). 10 μM antimycin A was used for baseline collection (measuring non-mitochondrial respiration). The OCR was normalized to protein content of each well.