Fig. 1

The process of four-color staining flow cytometry using monoclonal antibodies. The BMCs were collected and washed twice with FACS buffer. Cells were incubated with four antibodies against cell surface antigens, including CD45, CD34, CD38, and CD133 on ice for 30 min. a, b The live BMCs were collected, and SSC^low and CD45^dim cells were gated. c, d The BMCs were incubated with three types of combinations of monoclonal antibodies (mAbs) on ice for 30 min such as isotype control 1 (mouse anti-human CD45-FITC, mouse IgG-PE, mouse IgG-PE CY5 and mouse IgG-APC), isotype control 2 (mouse anti-human CD45-FITC, mouse anti-human CD34-PE, mouse anti-human CD38-PE CY5, and mouse IgG-APC), and sample (mouse anti-human CD45-FITC, mouse anti-human CD34-PE, mouse anti-human CD38-PE CY5, and mouse human CD133-APC). Cells were then washed twice with FACS buffer and analyzed using the FACSCalibur flow cytometer and CellQuest Pro software (BD Bioscience). Finally, the levels of CD45^dimCD34⁺CD38⁻CD133⁺ cells, CD133 positive cells among the R1, R2, R3-gated cells were measured and the results were expressed as percentage change from the baseline conditions including isotype control 2. The filled histogram represents the isotype control 2, and the empty histogram represents CD45^dimCD34⁺CD38⁻CD133⁺ cells