Figure 7

Responsiveness of neutrophil-like X-CGD PLB-985 cells and their parental counterparts. (A) Granulocytic (day 5) X-CGD PLB-985 cells (grey bars) or parental controls (open bars) were incubated for 30 min in the presence or absence of 100 ng/ml LPS, 100 U/ml TNFα, or 25 nM PMA (as a positive control), prior to the determination of superoxide generation by the cytochrome c reduction assay. Mean ± s.e.m. from at least 3 independent experiments. (B) Granulocytic (day 5) X-CGD PLB-985 cells (right panel) or parental controls (left panel) were incubated for 15 min at 37°C in the absence ("-") or presence of 100 ng/ml LPS, 100 U/ml TNFα, or 100 U/ml IFNγ, and nuclear extracts were prepared and analyzed in EMSA using a NF-κB or hSIE oligonucleotide probe. This experiment is representative of three. (C) Granulocytic (day 5) X-CGD PLB-985 cells (grey bars) or parental controls (open bars) were nucleofected with luciferase constructs driven by 5 repeated NF-κB elements ("pNFκB-Luc"), or by the IL-8 promoter ("pIL-8-Luc"), and cultured for 6 h at 37°C in the presence or absence of 100 ng/ml LPS or 100 U/ml TNFα, prior to cell lysis and luciferase activity measurement. Mean ± s.e.m. from 3 independent experiments.