Fig. 5

In vitro intervening F. nucleatum -induced tumor proliferation with BSA-Cu SAN on HCT116 cells. a CLSM images of HCT116 cells incubated with BSA-Cu-FITC containing 45 μg/mL Cu at different times. b Uptake of BSA-Cu SAN at different culture times analyzed by flow cytometry. (c) Relative contents of GSH in HCT116 cells treated with different concentrations of BSA-Cu SAN for 4 h. d Fluorescence images of HCT116 cells stained with DCFH-DA after various treatments with H₂O₂ only, BSA-Cu SAN only, and BSA-Cu + H₂O₂ for ROS detection. e Cell viability of NCM460 cells and f HK-2 cells incubated with different concentrations of BSA-Cu SAN for 24 h at pH 7.4. g Cell proliferation assay using CCK8. HCT116 cells were cultured with or without F. nucleatum (Fn) at different times. h Cell viability of HCT116 cells incubated with different concentrations of BSA-Cu SAN with or without 100 μM H₂O₂ at pH 6.5 for 24 h. i Western blot results of LC3 expression in HCT116 cells after various treatments (control, H₂O₂, Fn + H₂O₂, Fn + BSA-Cu, Fn + BSA-Cu + H₂O₂). j Relative expression of LC3 II in (i). k CLSM images of HCT116 cells stained with JC-1 after various treatments to detect the changes in mitochondrial membrane potential. Red fluorescence represents aggregates, while green fluorescence represents monomers. l Cell apoptosis results via flow cytometry analyses of HCT116 cells treated with BSA-Cu SAN with or without H₂O₂ . Scale bars = 50 μm. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001