Cellular Uptake and Colocalization of Materials

The 4T1 cells (mouse breast carcinoma cells) were purchased from the cell bank of the Chinese Academy of Sciences (Beijing, China) and maintained in RPMI 1640 (Gibco) medium containing 10% FBS (Gibco). The cells were seeded in six-well plates at 2.0×105/well, and cultured at 37°C and 5% CO2 for 24 h until they adhered to the bottom of the plate. The ICG, PHSA-ICG, and PHSA-ICG-TAT (10 μM) were added to the cells, respectively, and the cell suspension was collected after 0, 1, 4, and 8 h. The cell suspension was filtered with a 70 μM filter, and the cells were resuspended in 200 μL of PBS in a 1.5 mL Eppendorf tube. Monochromatic flow fluorescence analysis was carried out at 640 nm excitation and 780 nm emission to determine the cell uptake of materials. The above cells cultured for 24 h were stained with Hoechst staining solution for 15 min and observed using a fluorescence microscope (DP80; Olympus, Tokyo, Japan) for the colocalization of materials.

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