Pe conjugated anti b220

Peripheral blood was obtained at day 14 post-irradiation as described above. Red blood cells (RBCs) were lysed with 0.15 M NH₄Cl, followed by blocking with staining buffer (Hank's buffered salt solution plus 2% fetal bovine serum) containing F_c-block for 10 min before staining with antibodies. For analysis of differentiated cells, the cells were stained with PE-conjugated anti-CD11b (eBioscience) and PE-conjugated anti-Gr-1 (for myeloid cells) (eBioscience), APC-conjugated anti-Thy1.2 (for T cells) (eBioscience) or PE-conjugated anti-B220 (for B cells) (eBioscience) for 30 min on ice, followed by incubation with propidium iodide for 5 min on ice. Stained cells were washed with staining buffer and resuspended in 0.1 ml of staining buffer. For each sample, a minimum of 2×10⁵ cells were analyzed on a flow cytometer (LSR II, Becton Dickinson), and the data were analyzed with Cell Quest software (Becton Dickinson) after gating on viable cells. The percentage of B, T and myeloid cells in each mouse were calculated by multiplying the absolute numbers with the total numbers of WBCs harvested from each mouse per 0.1 ml of blood.