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Oxygen consumption rate Seahorse assay

SA Stephanie J. Alexopoulos
SC Sing-Young Chen
AB Amanda E. Brandon
JS Joseph M. Salamoun
FB Frances L. Byrne
CG Christopher J. Garcia
MB Martina Beretta
EO Ellen M. Olzomer
DS Divya P. Shah
AP Ashleigh M. Philp
SH Stefan R. Hargett
RL Robert T. Lawrence
BL Brendan Lee
JS James Sligar
PC Pascal Carrive
ST Simon P. Tucker
AP Andrew Philp
CL Carolin Lackner
NT Nigel Turner
GC Gregory J. Cooney
WS Webster L. Santos
KH Kyle L. Hoehn
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OCR was measured using an Agilent Seahorse XFe96 Analyzer (Agilent Technologies, Santa Clara, CA). NMuLi cells (American Type Culture Collection, Manassas, VA) were seeded in a 96-well seahorse plates at a density of 2.0 × 104 cells/well and were allowed to adhere overnight. Prior to the assay, media was changed to unbuffered DMEM containing pyruvate and glutamine (Gibco 12800-017) pH = 7.4 at 37 °C and the cells were equilibrated for 1 h at 37 °C without CO2. Compounds were injected during the assay and OCR was measured using 2 min measurement periods. Cells were treated with a single drug concentration. Three independent experiments were conducted with two to four replicates per treatment condition per experiment.

Copyright and License information: The Author(s) ©2020
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