2.9. Cell Cycle Analysis by Flow Cytometry

PC3-KD cells were incubated with ICRF-193 (2 μg/mL), HA-CdtB-NPs (50 nM, 100 nM, and 200 nM), and HA-NPs alone for 48 h. The cells were washed with PBS and centrifuged for 1000 rpm at room temperature for 5 min. Subsequently, the prepared cells were resuspended in hypotonic buffer (0.2 mg/mL RNase A, 20μg/mL propidium iodide, and 0.1% Triton X-100). Analysis of cell cycle was performed by flow cytometry (Becton Dickinson, San Diego, CA, USA).