Figure 4.

Analysis of the expression of NADPH oxidase in G6pc3^−/− macrophages. The CD11b-enriched macrophages used for function analysis were isolated from 6- to 8-week-old wild-type (+/+) and G6pc3^−/− (−/−) littermates. (A) Quantification of gp91^phox, p22^phox, and p47^phox mRNA in macrophages by real-time RT-PCR. Expression is normalized to β-actin and measured relative to 1 wild-type mouse arbitrarily defined as 1. Data represent the mean ± SEM of 4 independent experiments. (B) Western blot analysis of macrophage protein extracts with the use of Abs against gp91^phox, p22^phox, p47^phox, or β-actin. Data from 2 pairs of littermates are shown, and each lane contains 50 μg of protein. The relative protein levels of gp91^phox, p22^phox, and p47^phox were quantified by densitometry of 4 separate pairs of Western blots. The measurements are relative to β-actin. (C) Quantitative flow cytometric analysis of membrane-bound p47^phox in macrophages. The gray tracing represents the fluorescence background. Data represent the mean ± SEM of 4 independent experiments. **P < .005 and *P < .05.