Anti human cd71 pe

Assessment of cell surface markers expression by FACS was performed using a FACSCalibur or an Accuri C6 flow cytometer (BD Biosciences). Antibodies used are anti-SSEA4-PE (Miltenyi Biotec), anti-TRA-1-60-PE (Miltenyi Biotec), anti-SSEA4-AF647 (Biolegend), anti-TRA-1-60-FITC (BD Biosciences) and anti-human CD71-PE (Biolegend). FACS data were analyzed using FlowJo software (Tree Star).

CD34⁺ CML cells were stained with 1 μM CellTrace Violet (CellTrace Violet Cell Proliferation Kit, Life Technologies) in PBS for 30 min at 37 °C. The reaction was quenched by adding cell culture medium containing 10% FBS. Cells were then washed and re-suspended in SFM supplemented with PGF cocktail and treated as indicated in figure legends. After 3 or 6 days, cells were stained with anti-human CD34-APC (BD Biosciences), anti-human CD38-PerCP (BioLegend) and anti-human CD133-PE (Miltenyi Biotec). For detection of differentiation markers, cells were stained with anti-human CD71-PE, anti-human CD11b-PE-Cy7 and anti-human CD14-APC-Cy7 (all from BioLegend) followed by flow cytometry analysis (FACSVerse^TM Flow Cytometer, BD Biosciences). Data analysis was performed using FlowJo 7.6.5 software.