Figure 6.

gp91 expression rescues superoxide production after retargeting to heterochromatin. Superoxide production was measured in the transduced and electroporated X-CGD PLB cells. (a) Superoxide production was measured by the continuous cytochrome C reduction assay. (b) Percentage of NBT formazan-positive cells are depicted after reduction of NBT as measure for superoxide production. Mean ± SD of three measurements are shown. (c) Monoclonals were grown from the LV_gp91-transduced CBX1-LEDGF_325-530(D366N)-electroporated X-CGD PLB cells. MFI of gp91 expression per VCN. The MFI determined from the different gp91-expressing monoclonals was divided by the VCN of the respective monoclonal. (d) Superoxide production was measured by the continuous cytochrome C reduction assay for all different monoclonals and normalized to VCNs. CBX1, heterochromatin protein 1β eGFP, enhanced green fluorescent protein; gp91, glycoprotein 91; LEDGF/p75, lens epithelium-derived growth factor; MFI, mean fluorescence intensity; NBT, nitroblue tetrazolium; VCN, vector copy number; WT, wild-type; X-CGD, X-linked chronic granulomatous disease.