FIGURE 1.

Phagosomal reductive and oxidative processes in GILT^−/− BMMØs. BMMØs derived from WT and GILT^−/− mice were incubated for 18 h in the presence/absence of IFNγ. A, total mRNA levels of GILT in unactivated and IFNγ-activated BMMØs were determined by QPCR. Averaged relative mRNA levels from four independent QPCR experiments are shown. Relative expression was expressed as mRNA levels relative to 18 S and presented relative to unactivated BMMØs. Error bars denote S.E. *, p < 0.05. 10 min before addition and subsequent phagocytosis of experimental particles, BMMØs were treated with the NOX2 inhibitor DPI (0.5 μm) where indicated. B, C, and D, phagosomal disulfide reduction of the BODIPY FL l-cystine substrate conjugated to IgG-opsonized dextran-coated experimental particles. E and F, phagosomal oxidation of the OxyBURST Green H₂HFF BSA substrate conjugated to IgG-opsonized experimental particles. G, extracellular H₂O₂ production relative to unactivated/untreated WT control after phagocytosis of serum opsonized zymosan particles as measured by oxidation of the Amplex UltraRed reagent. B, C, and E, representative real-time traces. Relative fluorescence units (RFU) values are proportional to the degree of substrate reduction/oxidation. D and F, averaged rates relative to unactivated/untreated WT controls of four independent experiments are shown for each phagosomal measurement. Average relative rates of disulfide reduction were calculated between 40 and 60 min after particle internalization. Relatives rate of OxyBURST Green H₂HFF BSA oxidation were calculated between 30 and 80 min after particle internalization. A, D, F, and G, error bars denote S.E. of four independent experiments. *, p < 0.05.