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. 2018 Jun 25;182(4):567–578. doi: 10.1111/bjh.15441

Figure 1.

Figure 1

Effect of xanthine oxidase and hypoxanthine (XO/HO) mixtures on accumulation of reactive oxygen species (ROS) and phosphatidylserine (PS) exposure in red cells from patients with sickle cell anaemia (SCA). (A) Red cells were pre‐loaded with CM‐H2DCF‐DA (100 μmol/l) to measure ROS levels [as median fluorescence intensity (MFI)] or treated with the same final concentration of dimethyl sulphoxide [DMSO] (control) before incubation with hypoxanthine (HO, 2 mmol/l) and xanthine oxidase (XO, 0–0·1 U/ml) mixtures for 30 min at 37°C (n = 2). (B) Red cells were permeabilised to Ca2+ with the ionophore bromo‐A23187 (6 μmol/l), and incubated in different free [Ca2+]os, maintained using Ca2+ / 2 mmol/l EGTA mixtures, at 0·5% haematocrit for 30 min at 37°C in the absence (filled circles) or presence (open circles) of HO (2 mmol/l)/XO (0·015 U/ml) mixtures after which externalised PS was labelled with LA‐FITC, n = 8. PS exposure was normalised to that of control red cells at 1 μmol/l free [Ca2+]o (31·1 ± 3·9% of total red cells). Symbols represent means ± SEM for red cells from n different individuals. *P < 0·05; **P < 0·005.