Intermittent Hypoxia Affects the Spontaneous Differentiation In Vitro of Human Neutrophils into Long-Lived Giant Phagocytes

Figure 5

Effects of hypoxia on gp91-phox and p22-phox expression in giant phagocytes (Gϕ). Freshly isolated PMN were exposed for 24 h to intermittent hypoxia (IH, 56 cycles), sustained hypoxia (SH), or normoxia (N) and then cultured at normoxia for additional six days. For double immunofluorescence staining fixed cytospins were stained with rabbit anti-gp91-phox and mouse anti-p22-phox primary Abs (1/100) or the corresponding isotype controls (rabbit IgG and mouse IgG2) followed by 1/400 CF 488A goat anti-rabbit IgG (green) and CF 647 goat anti-mouse IgG (red) staining. Nuclei were stained with DAPI (blue). Representative data out of 3 independent experiments.