For apoptosis analysis, ESCC cells transfected with siRNA were harvested after 48 h. According to manufacturer's instructions, the transfected cells were treated with fluorescein isothiocyanate (FITC)-Annexin V and propidium iodide (PI) for 30 min at room temperature off from the light. Subsequently, the cell apoptosis were analyzed by flow cytometry (FACScan; BD Biosciences, Shanghai, China).

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