Figure 3.

FAC-induced defective monocyte to macrophage differentiation is partially rescued by the antioxidant NAC. (A) Surface expression of CD206 on macrophages differentiated with M-CSF for 3 days in the presence of FAC (4 µg Fe/mL), deferoxamine mesylate salt (DFO, 87.5 μM), NAC (4mM), or vehicle 0.4% DMSO. (B) Phagocytic capacity of macrophages differentiated with M-CSF in the presence of FAC (4 µg Fe/mL), DFO (87.5μM), NAC (4mM), or vehicle 0.4% DMSO. Data are shown as the increment in fluorescence intensity of engulfed pHrodo E. coli bioparticles between cells incubated at 37°C and at 4°C. Each dot represents an individual donor (n = 7-8). Data shown are pooled from two experiments. One-way ANOVA with Dunnett’s multiple comparisons test was performed. *p < 0.05, **p < 0.01. (C) Quantification of mitochondrial mass in monocytes differentiated with M-CSF in the presence or absence of FAC (4µg Fe/mL) for 6h, 1 day and 5 days, in comparison to cells differentiated with M-CSF alone and presented as Mitotracker gMFI (n = 3-6). (D) Quantification of mtROS production in monocytes differentiated with M-CSF and treated with FAC (4µg Fe/mL) or a combination of FAC (4µg Fe/mL) and NAC (4mM) for 24h (n = 6) and 5 days (n = 4). Data are presented as MitoSOX gMFI. Each symbol represents an individual donor. One-way ANOVA with Dunnett’s multiple comparisons test was performed. *p < 0.05.