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. 2018 Jun 21;9:1404. doi: 10.3389/fimmu.2018.01404

Figure 3.

Figure 3

Effect of priming agents on eosinophil and neutrophil oxygen consumption rates. (A) Kinetic oxygen consumption rates (OCR) of eosinophils following injection of PMA (closed circles, 200 nM) at 15 min or PAF (open squares, 5 µM) at 45 min. Eosinophils stimulated with PAF were exposed to IL-5 (10 ng/ml) for 30 min prior to injection of PAF. Closed squares represent the media only injections. Data represent the mean ± SEM of three independent experiments. (B) Kinetic oxygen consumption rates (OCR) of eosinophils following injection of IL-5 alone at 15 min followed by fMLP injection (open squares, 100 nM) at 45 min. Closed squares represent the media only injections. Data represent the mean ± SEM of seven independent experiments (*p < 0.05 compared with media alone using a Student’s t-test). (C) Kinetic OCR responses of neutrophils following injection of PMA (closed circles, 200 nM) at 15 min or fMLP (open squares, 100 nM) at 50 min. Neutrophils stimulated with fMLP were injected with GM-CSF (10 ng/ml) for 35 min prior to fMLP treatment. Black squares represent the media only injections. Data represent the mean ± SEM of three independent experiments. (D) Comparison of the peak height OCR by eosinophils (white bars) and peak height OCR by neutrophils (black bars). Treatments were performed as described in (A,C). Data represent the mean ± SEM of four independent experiments for GM-CSF/fMLP stimulation, five independent experiments for PAF/fMLP, and ≥4 independent experiments for PMA (*p < 0.05 compared with media alone using a Student’s t-test). Ns indicates a non-significant difference. (E) Comparison of eosinophil peak height OCR following PMA (200 nM) injection under normoxia or hypoxia. Data represent the mean ± SEM of ≥4 independent experiments under normoxia (*p < 0.05 compared with media alone using a Student’s t-test) and mean ± SD of two independent experiments under hypoxia. (F) Comparison of neutrophil peak height OCR following PMA (200 nM) injection under normoxia or hypoxia. Data represent the mean ± SEM of ≥4 independent experiments under normoxia (*p < 0.05 compared with media alone using a Student’s t-test) and mean ± SD of two independent experiments under hypoxia. Eosinophil and neutrophil OCR responses were measured simultaneously on the same assay plate (D,E).