Fresh healthy donor human PBMCs were incubated with CellROX (Thermo Fisher Scientific), NAO (Thermo Fisher Scientific), or 2-NBDG (Carbosynth) according to the manufacturer’s specifications. For all three conditions, cells were stained during the efflux incubation for MitoTracker and otherwise prepared for flow cytometry using the protocol detailed above with a few exceptions. 2-NBDG and NAO panels required substitution of MitoTracker Orange (Thermo Fisher Scientific). The fluorochrome-conjugated primary antibody panel was as follows: CD3 (HIT3a, BioLegend), CD19 (SJ25C1, BioLegend), CD27 (M-T271, BD Biosciences), IgD (IA6-2, BioLegend), and CD10 (HI10a, BioLegend). Compensation for the functional dyes required the use of Ramos cells (ATCC), incubated with CellROX, 2-NBDG, or NAO, respectively.

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