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Seahorse analysis

MN Marie-Julie Nokin
FD Florence Durieux
JB Justine Bellier
OP Olivier Peulen
KU Koji Uchida
DS David A. Spiegel
JC James R. Cochrane
CH Craig A. Hutton
VC Vincent Castronovo
AB Akeila Bellahcène
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Oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) of U87-MG, U251, MDA-MB-231 and MCF7 cells cultured in low and high glucose medium were determined using Seahorse XFp bioenergetic analyzer (Agilent) according to manufacturer’s recommendations. Briefly, cells were plated in Seahorse microplates (15000, 5000, 15000 and 20000 cells/well for U87-MG, U251, MDA-MB-231 and MCF7 cells, respectively). Medium was replaced by DMEM without glucose supplemented with 2 mM glutamine 1 h before the measurements. D-glucose was added to a final concentration of 10 mM and OCR and ECAR were measured. Data were normalized to protein quantity.

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