4T1 cells (1 × 105 cells per well) were seeded in 12-well plates and cultured for 24 hours. Then, cells were incubated with free CPT, CSSP NPs, or CP NPs (10 μM, CPT equivalent) for 12 or 24 hours (n = 3). After incubation, the cells were washed, fixed with 4% paraformaldehyde, and counterstained with Hoechst 33342. The round coverslips were observed by TCS SP2/AOBS confocal laser scanning microscopy (CLSM) (Leica, Germany). For flow cytometry analysis, after incubation with different formulations, cells were washed, digested, and suspended in PBS. The intracellular fluorescence intensity was determined by a FACSCalibur flow cytometer (BD, USA).

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