Table 1.
Functional analysis of wild-type and transgenic COS7 cell lines in cell-free NADPH oxidase assays
| Source of membranes | O2− production*, RLU | O2− production†, mmol/min/mg protein |
|---|---|---|
| COS7 WT | 12,711 ± 27,000 (n = 3) | 0.028 ± 0.9 (n = 6) |
| COS7 gp91 | 11,111 ± 22,555 (n = 3) | 0.13 ± 0.22 (n = 6) |
| COS7 p22 | 67,867 ± 7,669 (n = 3) | −0.06 ± 0.03 (n = 6) |
| COS7 gp91/p22 | 295,034 ± 81,407 (n = 3) | 17.0 ± 0.37 (n = 3) |
| Neutrophils | 538,990 ± 52,888 (n = 3) | 88.5 ± 13.3 (n = 3) |
Cell-free assays were performed as described in Materials and Methods by using neutrophil or COS7 cell membranes and neutrophil cytosol as a source of cytosolic oxidase components. O2− production was monitored by using a chemiluminescence detection system, and the results are expressed as mean ± SEM Tiron-inhibitable relative luminescence units (RLU).
Cell-free assays were performed as described in Materials and Methods by using partially purified neutrophil or COS7 cell membranes and neutrophil cytosol as a source of cytosolic oxidase components. O2− production was monitored by using a cytochrome c detection system, and the results are expressed as mean ± SEM SOD-inhibitable O2− production.