LSCs apoptosis assay in AML samples

The effect of alantolactone on LSC cells was analyzed by flow cytometry. The primary AML mononuclear cells were obtained from AML patients. The mononuclear cells which was isolated from primary AML patients were seeded in 24-well plates (1 × 10⁶ cells/well), and medium volume was 1 mL. Then, cells were treated with various concentrations of alantolactone. After 18-h treatment, the cells were re-suspended with PBS and stained with CD34-APC and CD38-PE.cy7 for 30 min, and then the cells were re-suspended with 1× binding buffer which contained 5 μL Annexin-V-FITC and 5 μL PI. Samples were analyzed by flow cytometry in 1 h.