Figure 1.

Decreased reactive oxygen species (ROS) production after Toll‐like receptor 7 ( TLR7) or TLR9 stimulation in NADPH oxidase‐deficient B‐cell lines. (a) AR, a B‐cell line derived from a patient with mutations in both paternal and maternal alleles of p40^phox and ARp40, the AR cell reconstituted with wild‐type p40^phox, were lysed and the lysates were immunoblotted to detect p40^phox and GAPDH proteins. (b) Cell lysates from KS, a B‐cell line derived from a patient lacking functional p47^phox and KSp47, the KS B‐cell line reconstituted with p47^phox, were immunoblotted to detect p47^phox and GAPDH proteins. (c,d) ROS production by B cells was monitored after a 30‐min treatment with diphenyleneiodonium (DPI), R848, CpG and/or PMA and a 10‐min incubation with 2′7′‐dichlorodihydrofluorescein diacetate (H₂ DCFDA) as described in the Materials and methods. Mean fluorescence intensity (MFI) was detected by flow cytometry and plotted as a ratio to the MFI of cells treated with H₂ DCFDA alone which was set to one (dashed line). Data shown in (c) and (d) represent one of three independent experiments.