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Ascorbate measurement For neutrophil ascorbate measurement, 10 l of the extraction solvent (80% methanol containing 10 mM EDTA and 50 pmoles 13 C-ascorbate/200,000 cells as an internal standard) was added to 10 L neutrophil extract, vortexed for 10, centrifuged at 21,000 g for 15 min at 4 C, the supernatant was diluted using 30 L of 0.03% formic acid in water and analyzed using an AB Sciex 6500+ QTrap mass spectrometer coupled to a Shimadzu UHPLC system operating in negative mode to detect the transitions 175/115 (ascorbate) and 176/116 ( 13 C-ascorbate)

Almanza A, Carlesso A, Chintha C, Creedican S, Doultsinos D, Leuzzi B, Lus A, McCarthy N, Montibeller L, More S, Papaioannou A, Pschel F, Sassano ML, Skoko J, Agostinis P, de Belleroche J, Eriksson LA, Fulda S, Gorman AM, Healy S et al (2019) Endoplasmic reticulum stress signalingFrom basic mechanisms to clinical applications

383,384 The secondary mtDNA depletion may exacerbate OXPHOS disturbance, increasing the production of mtROS

Cellular effects measured over months

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