p22phox is involved in Mo-DC differentiation and maturation. (A-C) Freshly isolated Mos from 2 p22phox CGD patients (experiments repeated twice) were treated with or without NAC or DPI at indicated concentrations 15 minutes prior to GM-CSF/IL-4 differentiation stimuli or LPS maturation stimuli. Differentiation was carried out for 6 days and LPS maturation for 2 more days. On day 7 (immature Mo-DC) or day 9 (mature Mo-DC), cells were harvested and prepared for flow cytometry analysis after incubation with CD86 PE/CD209 PerCP-Cy5.5/CD83 APC cocktail antibodies for 20 minutes in the dark. Data are presented as MFI. (D) Mos were transfected with 200 nM scrambled or p22phox siRNA. Five days later, cells were harvested and prepared for flow cytometry analysis after incubation with CD86 PE/CD209 PerCP-Cy5.5/CD83 APC cocktail antibodies for 20 minutes in the dark. Data are presented as MFI of CD209 and are expressed as mean ± SEM of n = 4. (E) Representative western blot analysis of Mo-DC siRNA transfection and (F) relative quantification (n = 4, arbitrary units). One-way ANOVA with Tukey test. *P < .05, ***P < .001.