Figure 2
From: IVIG regulates the survival of human but not mouse neutrophils
Confirmation of selective survival regulatory capacity of IVIG in human but not mouse neutrophils by different death assays. Features of cell death assessed in GM-CSF primed or unprimed human (PMNH) or C57BL/6 mouse (PMNBL/6) neutrophils following treatment with IVIG by flow cytometry (A–C) or light microscopy (D). (A) Annexin V-FITC/PI staining assay. Quantitative analysis is indicated as the percentage representative of each quadrant. (B) DNA-fragmentation assay. Quantitative analysis of gated subdiploid DNA is indicated. (C) Assessment of mitochondrial potential (Δψm). The mitochondrial uncoupler FCCP was used as a positive control. Results of 5- (C) or 15-hour cultures (A,B,D) are shown. (D) Morphological characterization. Cells were stained with Giemsa–May–Grünwald (Diff-Quik) (original magnification X1000). Continuous arrows indicate condensed nuclei, dashed arrows vacuoles. The lower panel represents the statistical analysis of the normal, apoptotic or vacuolated phenotypes. Data are representative for at least 3 independent experiments. Bars show mean ± SEM. *p < 0.05, **p < 0.01, one way-ANOVA followed by Dunnet’s post hoc test. Δψm = mitochondrial potential.
