Pe conjugated mouse anti human cd133 clone ac133

After 48 h of vehicle or CDDP treatments, A549 cells were harvested, washed and stained with PE-conjugated mouse anti-human CD133 (clone AC133, Miltenyi Biotech, Paris, France) or PE-conjugated mouse control isotype (Miltenyi Biotech) in PBS plus 2 mM EDTA and 0.5% BSA on ice. In all conditions, doublet discrimination was performed to gate only single cells and nonviable cells exclusion was assessed using 7-amino-actinomycin D probe (7-AAD). CD133⁺ A549 subpopulations were sorted by FACS (Fluorescence-Activated Cell Sorting). Cell analysis and aseptic cell sorting was performed with a FACSAria II flow cytometer (Becton-Dickinson, Franklin Lakes, NJ, USA). FACS data for three independent experiments were analyzed using the FlowJo V10 software (TreeStar, Inc., Ashland, OR, USA).

CD34+ cells from NN or LN were washed in PBS (1X), and stained with monoclonal antibodies for FACS analysis: APC-conjugated mouse anti-human CD34 (clone AC136, Miltenyi Biotec, Auburn, CA, USA), PE-conjugated mouse anti-human CD133 (clone AC133, Miltenyi Biotec, Auburn, CA, USA), FITC-conjugated mouse anti-human CD38 (clone IB6, Miltenyi Biotec, Auburn, CA, USA), and PE-conjugated mouse anti-human CD117 (clone AC126, Miltenyi Biotec, Auburn, CA, USA), and analyzed as described above.