Evaluating Cell Surface Markers by Flow Cytometry

The cell surface markers of differentiation were evaluated by flow cytometry as described in our previous study [16 (link)]. AML cell lines or human primary AML cells were cultured in 6-well plates at a density of 1 × 10⁶ cells per well and treated with different compounds for 72 h. Subsequently, treated cells were resuspended in phosphate-buffered saline containing 1 mM EDTA and 2% FBS, and incubated with FITC-conjugated anti-CD11b or PE-conjugated anti-CD14 (BioLegend) for 30 min in the dark at room temperature. Flow cytometer data were collected by BD FACS Caliber (BD Biosciences, NJ, USA) and analyzed through FlowJo (BD Biosciences, NJ, USA).