Cytofluorimetric analysis of fas and Annexin-V on different bone marrow populations. Bone marrow cells (2 × 105/mL) were cultured 7 to 18 hours with rGM-CSF, rIL-3, and rIL-5 in the presence of 2.4G2 (25 μg/mL). Cells were stained with Gr-1 (PE) as a marker of the granulocyte population and with IgM (Cy5) as a marker of the lymphocyte population; Annexin-V (FITC) or fas (FITC) were added, and PI-positive cells were excluded in all experiments. A granulocyte gate and a lymphocyte gate were made based on the orthogonal and forward scatter. Fas expression was studied by comparing isotype-matched antibody (broken line), BM cells cultured in the presence of cytokines (thin line), and cells cultured with cytokines plus 2.4G2 (thick line). (A) Apoptotic analysis of the granulocytic cells. (B) Apoptotic analysis on the lymphocyte gate. The figure shows a representative experiment from 5 with similar results.
