Fig. 4

HTS NPs-mediated ROS induce lung cancer cell death. (A) MTT assay of H460 cells treated with the HTS NPs and in combination with or without Z-VAD, Fer-1 and CQ (n = 3). (B-C) Annexin V-FITC/PI staining analysis of apoptosis by flow cytometry (n = 3). (D) Western blot analysis of apoptotic markers for H460 cells treated with TPGS, STZ, TS NPs and HTS NPs. (E-F) ROS generation in H460 cells after different treatments using fluorescence imaging and flow cytometry. Scale bar: 100 μm. (G) MTT assay, colony formation assay (H-I) and LDH assay (J) of H460 cells were treated with HTS NPs with or without NAC (2 µM) treatment (n = 3). (K-L) Fluorescence imaging of AM/PI staining assay and quantification analysis (L) in lung cancer cells after different treatments with HTS NPs with or without NAC (2 µM), scale bar: 100 μm. (M) Immunoblot analysis of apoptotic markers for H460 cells treated HTS NPs with or without NAC treatment. (N) Annexin V-FITC/PI staining analysis of apoptosis H460 cells by flow cytometry. The error bars represent the means ± SD (n = 3, ***P < 0.001)