Cell Cycle and Differentiation Analysis

Cell-cycle profile was analyzed using flow cytometry. Briefly, 2 × 10⁶ cells were washed with PBS, fixed in 80% ethanol and resuspended in 50μg/ml propidium iodide (Sigma) and 125U/ml RNase A (Sigma). For cell cycle profile, DNA content was analyzed by flow cytometry using FACScan and Cell Quest Software (Becton Dickinson, USA). For detection of cell differentiation antigen cd11b and cd114, 1 × 10⁶ cells were washed twice with PBS, incubated with PE-conjugated cd11b or cd114 antibody or PE-conjugated IgG isotype control antibody at 4°C for 30 min and analysed by flow cytometry using FACScan and Cell Quest Software (Becton Dickinson, USA) [12 (link), 13 (link), 29 (link)].