, Glycolysis and glycolytic capacity values were calculated by subtraction of ECAR value after treatment of cells with 2-deoxyglucose (which corresponds to non-glycolytic acidification) from ECAR values in cells treated with glucose or with oligomycin, respectively. Measurement of fatty acid oxidation Fatty acid oxidation was measured as described previously, OCR value after treatment of cells with rotenone and antimycin A (which corresponds to non-mitochondrial respiration) from OCR values in cells treated with glucose or with oligomycin and FCCP, respectively, 2012.
, Metabolomics Metabolite extraction of GM-DC was performed on 2.5 million per well using 70°C aqueous 70%
, At collection, cells were placed immediately on ice, the media was removed and cells were washed three times with ice-cold PBS to remove residual media. Intracellular metabolites were extracted twice with hot ethanol using 10µM norvaline as an internal control. For LCMS, samples where dried under nitrogen flow and reconstituted in a milliQ water/acetonitrile (1:1) mixture for injection using a UPLC Acquity (Waters) separation system coupled with a Xevo G2 ToF (Waters) as described (Paglia et al., 2012) with slight modification. Compounds were ionized using an electrospray ionization source in negative mode, 2015.
, Raw data was converted to netCDF format using Chemstation (Agilent), before processing in Matlab R2014b (Mathworks, Inc.) using PARADISe software as described, Compound identification was performed using both retention time of authentic standards and accurate mass with an accepted deviation of 0.005 Da, 2017.