Figure 3

Annexin V/PI analysis in induced sputum. (a) Horizontal axis represents side scatter (SS) and vertical axis represents forward scatter (FS) (linear scale). Gated area, containing granulocytes, represented by area marked M (b) Horizontal axis represents intensity of staining for Av (Annexin V) (logarithmic scale) and vertical axis intensity of staining for PI (logarithmic scale). Typical dot plot representing populations of early apoptotic (AV⁺/PI^-), secondary necrotic (AV⁺/PI⁺), and necrotic granulocytes (AV⁺/PI^-) (c) Analysis of neutrophil apoptosis in sputum using the Annexin V/Propidium iodide assay. One way ANOVA showed a reduction in Av⁺/PI^- in healthy smokers (HS) and COPD subjects compared to non-smokers (NS), p < 0.001. % AV⁺/PI^-: early apoptosis, % AV^+/PI⁺: later stage apoptosis/secondary necrosis:% AV^-/PI⁺: necrosis.