Apoptosis assay.

(A) Histogram plots from a representative experiment in which bone marrow neutrophils isolated from a heterozygous V72M NE mouse were cultured in the absence of G-CSF and harvested at the indicated times. Cells were stained for Gr-1, annexin V, and 7-AAD. Shown is annexin V and 7-AAD staining after gating for Gr-1⁺ (granulocytic) cells. The percentages of viable (annexin V⁻, 7-AAD⁻) cells within the Gr-1⁺ population were 91.8%, 64.5%, and 9.8% on days 0, 1, and 2, respectively. (B,C) Neutrophils isolated from the bone marrow of mice of each genotype (n = 3, each) were cultured in the absence (B) or presence (C) of 100 ng/mL G-CSF. The percent of viable cells is shown. Data represent mean ± SEM.