Neutrophil differentiation of X-CGD iPSC line iXC9 and normal control human iPSC line iPS(IMR90). Panels A-C show cytospin Giemsa stain, cytospin NBT assay, and DHR flow cytometric analysis, respectively, of neutrophils arising from differentiation of normal control iPSC line iPS(IMR90). Panels D-F are cytospin Giemsa stain, cytospin NBT assay, and DHR flow cytometric analysis, respectively, of neutrophils arising from differentiation of X-CGD iPSC line iXC9. Arrows in the cytospin Giemsa stain images (A,D) indicate some of the cells with mature neutrophil morphology (bar = 10 μm). Cytospin NBT assay shows blue-black formazan precipitates resulting from robust ROS production by PMA-activated neutrophils derived from normal control iPSCs (B), but lack of formazan precipitates indicating no ROS production by neutrophils derived from X-CGD iPSCs (E). The DHR dot plot images (y-axis side scatter; x-axis DHR fluorescence) demonstrate robust ROS production (high fluorescence) by 38.5% of PMA-activated cells representing mature neutrophils derived from normal control iPSCs (C), but no evidence of any significant ROS production by PMA-activated neutrophils derived from X-CGD iPSCs (F).