The apoptosis assay was performed after transfection using the Annexin V-fluorescein isothiocyanate Apoptosis Detection Kit I (BD Biosciences, San Jose, CA, USA) and assessed by flow cytometry. CD133 antibodies were obtained from eBioscience (CA, USA). OS cells were incubated with CD133 antibodies (1:1000) in PBS containing 1% bovine for 30 min at 4 °C. After washing with PBS, the cells were analyzed by flow cytometry (Cyto FLEX, Beckman, Germany). The results were analyzed using Cyt Expert.

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