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Oxygen consumption rate measurements.

GK Gyu-Lee Kim
TH Thomas A. Hooven
JN Javiera Norambuena
BL Barry Li
JB Jeffrey M. Boyd
JY Jason H. Yang
DP Dane Parker
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Bacterial respiratory activity was quantified using the Seahorse XFe96 extracellular flux analyzer (Agilent Technologies, Santa Clara, CA) as previously described (28, 57). Overnight cultures of S. aureus cells in LB were diluted 1:100 in M9 minimal medium with 1% Casamino Acids and grown to the mid-exponential phase at 37°C. Cultures were diluted to an OD600 of 0.05, and 200 μl of diluted cells was dispensed into each well of XFe cell culture microplates precoated with 100 ng/ml of poly-d-lysine in technical triplicate. CFU were plated for reach sample. Microplates were centrifuged for 10 min at 1,600 relative centrifugal force (RCF) to adhere cells to XFe microplates. Measurements were taken at 5-min intervals with a cycle of 2.5 min of measurement and 2.5 min of mixing, with measurements averaged over the first three cycles. OCR and ECAR values displayed are normalized by CFU and averages across n ≥ 3 biological replicates for each strain.

Copyright and License information:  ©2021 Kim et al.
This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.

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