Fig. 6
Anticancer efficacy of FeS2-based NPs in MNNG-HOS cells. Intracellular ROS level assessed with (A) the percentages of cells producing ROS and (B) the MFI of ROS detected by flow cytometry with DCFH-DA probe in MNNG-HOS cells after FeS2-based NPs treatment for 24 h. (C) Intracellular LPO level detected by flow cytometry with C11-BODIPY 581/591 probe in MNNG-HOS cells after FeS2-based NPs treatment for 24 h. (D) Quantitative MFI analysis of LPO recorded in panel (C). Loss of intracellular GSH assessed with (E) the rate of GSH/total glutathione and (F) the rate of GSH/GSSG in MNNG-HOS cells after FeS2-based NPs treatment for 24 h. (G) Western blot analysis of GPX4 expression in MNNG-HOS cells treated with different nano-formulations for 24 h with or without laser irradiation, and (H) GPX4 protein expression was quantified and normalized with β-actin. (I) Quantitative analysis of JC-1 monomers rate to assess the mitochondrial membrane potential of MNNG-HOS cells induced by FeS2-based NPs treatment for 24 h. (J) Quantitative analysis of MNNG-HOS cells apoptosis induced by FeS2-based NPs treatment for 24 h. (K) Cell viability of MNNG-HOS cells was analyzed by a CCK-8 assay after the treatment with FeS2-based NPs for 24 h. All data were expressed as the mean ± SD (n = 3. Statistics were done using one-way ANOVA with Tukey multi-comparisons. *p < 0.05, **p < 0.01, ***p < 0.001 and ns, no significance)