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. 2021 Jul 12;22(14):7474. doi: 10.3390/ijms22147474

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Mitochondrial function and glycolysis are impaired in HGPS fibroblasts. (A) Schematic representation of Seahorse XF Cell Mito stress test and calculated values are indicated. Oxygen consumption rates (OCR) (B) and extracellular acidification (ECAR) (E) were determined with a Seahorse XF96 Flux analyzer in basal and stimulated conditions (n = 3). Additional parameters like basal respiration (C), spare respiratory capacity (D), and basal ECAR levels (F) were calculated with Wave software v2.6.1.53 (Agilent Technologies, Santa Clara, CA, USA). (G) Cellular ATP levels were measured using a CellTiter-Glo luminescence ATP assay. (H) Intracellular ROS levels were determined by measuring oxidized dichlorofluorescein (DCF) levels using a DCFDA cellular ROS detection assay (n = 3). Graphs show mean ± SD. (* p < 0.05, ** p < 0.01, *** p < 0.001).