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. 2021 Sep 2;162(12):bqab188. doi: 10.1210/endocr/bqab188

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© The Author(s) 2021. Published by Oxford University Press on behalf of the Endocrine Society.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Mitochondria analysis and cellular adenosine 5′-triphosphate (ATP) production of orbital adipose tissue preadipocytes/fibroblasts (OAT-PFs) in adipogenic-medium (ADM)-adipogenesis compared with white adipose tissue (WAT). Confluent PFs from healthy OAT (OAT-H), Graves orbitopathy (OAT-GO), or WAT were cultured in complete medium (CM) or ADM for 4 days. A, DNA extracted from OAT (n = 3) and OAT-GO (n = 3) or WAT-PFs (n = 4) on day 4, relative quantitative polymerase chain reaction of cytochrome b (Cytob, mitochondrial DNA) to RPL13A (reference genomic DNA) was performed. B, Mitochondrial-cytochrome-oxidase (MtCO₂) antibody was analyzed by flow cytometry from confluent OAT (n = 4), OAT-GO (n = 4), and WAT-PFs (n = 7) at day 0 in CM or day 4 in CM and ADM, the percentage of flow cytometric (FL) intensity, positive mitochondrial-cytochrome-oxidase (MtCO₂) staining, is shown referenced to negative control. C, MTS assay (indicating the production of NAD(P)H and mitochondrial oxidative-phosphorylation (OXPHOS) activity (26)) was performed on PFs from OAT-H (n = 3) and WAT-PFs (n = 4) on day 4 in CM and ADM, 490 nm absorbance normalized by cell number. D, Total level of cellular ATP (luminescent ATP detection assay kit) was measured from OAT-H (n = 7) and WAT-PFs (n = 4) on day 4 in CM and ADM. Histograms = mean ± SEM of all samples studied. T test was used for statistical analysis. ***P less than or equal to .001; ****P less than or equal to .0005.