Quantitative analysis of p47phoxexpression by FACS analysis and immunoblotting. To determine p47phox expression, whole blood was permeabilized/fixed and then stained with anti-p47phox antibody. Neutrophils were gated using forward and right-angle light scatter. The level of p47phox expression on neutrophils is presented as the mean fluorescence intensity (MFI). (A) Relative histograms of a p47phox CGD patient, a p47phox CGD carrier, and a healthy volunteer. Summary data of the different populations are presented as scatter plots (B). Solid red circles in panels B, C, and D represent p47phox patients and carriers with non-ΔGT mutations in NCF1. (C) A subset of patients and carriers of p47phox CGD and healthy volunteers were analyzed for p47phox expression by immunoblotting. The data for of p47phox expression are presented as arbitrary units relative to the expression of β-actin. Neutrophil lysates from at least 2 healthy normal volunteers were analyzed on each gel, and the mean ratio of p47phox:β-actin for the normal samples was set to a value of 1.0. (D) The correlation obtained from parallel studies analyzing the expression of p47phox by both flow cytometry and immunoblotting. ****P < .0001 (1-way analysis of variance using Tukey’s multiple comparison).