H1299 cells (1 × 105) were seeded in 12-well plates, cultured for 48 hours, and transiently transfected with 1 µg of the indicated p53 plasmids. Then, the cells were treated with a Propidium Iodide (PI)/Fluorescein Isothiocyanate (FITC)-Annexin V Staining Kit (Vazyme, A211-01) and subjected to flow cytometry. A total of 106 cells were analyzed for each sample, and each condition was tested in three independent samples. FlowJo software was utilized to calculate the apoptotic cell rate.

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