Figure 4.

Quantification of ROS in CYBB-gene-corrected cell lines, as detected by DHR assay. (A) Monocytes and (B) macrophages from WT NHDF1 iPS cells (WT), a P47^Phox mutant iPS cell line (CGD1), a CYBB mutant iPS cell line (CGD2), the mixed pool CGD2.GC16A of gene edited iPS cells (GC16A), and the two gene-edited single-cell clones from CGD2.GC16A (C4 and E4) were stimulated with PMA or with PMA and fMLP, respectively, for 30 min in the presence of DHR, then immediately analyzed by flow cytometry (BD FACSCaliber). ROS generation is detected by oxidation of DHR into a fluorescent product and quantified as fold change (top right of each panel) in mean fluorescence intensity of DHR fluorescence (dark grey) compared with the unstimulated cells (light grey). Results shown are representative of two independent experiments.