Oxygen consumption rate (OCR) was measured with a Seahorse XFe24 analyzer (Agilent). ARVMs were seeded in 24-well Seahorse assay plates in complete medium and, after 2 h of cell attachment, the medium was replaced with XF basal medium supplemented with 10 mM glucose, 4 mM glutamine, and 1 mM sodium pyruvate (pH 7.4). Plates were incubated for an additional 1 h at 37 °C in a CO2-free incubator. The hydrated wells of the sensor cartridge were then loaded with 1 μM oligomycin (port A), 1 μM carbonyl cyanide 4- (trifluoromethoxy) phenylhydrazone (FCCP) (port B) and 1 μM antimycin A + 1 μM rotenone (port C). The data were analyzed using the Seahorse Wave software.
Copyright and License information: The Author(s) ©2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
Do you have any questions about this protocol?
Post your question to gather feedback from the community. We will also invite the authors of this
article to respond.