Following each treatment cycle, cells were harvested, washed twice with PBS, and fixed in 70% ethanol at − 20 °C for 16 h. The fixed cells were washed twice with PBS and stained with PI for 15 min at 37 °C. Analysis of cells with sub-G1, G0/G1, S, and G2/M DNA content was performed using 10,000 cells on an LSR Fortessa flow cytometer (BD Biosciences). Data were analysed using the FACSuite software (BD Biosciences).

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