Additional file 1 of Early diagnosis of breast cancer lung metastasis by nanoprobe-based luminescence imaging of the pre-metastatic niche

Additional file 1: Materials and Methods. Scheme S1. Synthesis of PPV. Fig. S1. Structural characterization of a luminescent material LCD. Fig. S2. Characterization of PPV. Fig. S3. The luminescence spectrum of luminol in the presence of hypochlorite. Fig. S4. TEM images of LAD NPs containing various contents of PPV. Fig. S5. Luminescence profiles of LAD NPs at pathophysiological concentrations of H2O2. Fig. S6. A sketch showing preparation of LCD nanoparticles (LCD NPs) by nanoprecipitation. Fig. S7. Comparison of luminescence intensities of LAD NPs based on three different batches in the presence of ClO-. Fig. S8. The effects of different inhibitors on cellular uptake of LAD NPs in neutrophils. Fig. S9. Dose-dependent luminescence of LAD NPs in peritoneal neutrophils. Fig. S10. Changes in the mean diameter of LAD NPs after incubation with PBS or neutrophil lysates. Fig. S11. Flow cytometric analysis of the distribution of LAD NPs in neutrophils in the lung tissue of mice at week 3 after i.v. inoculation of 4T1-GFP tumor cells. Fig. S12. Spectroscopy characterization of PGP-conjugated DSPE-PEG (DSPE-PEG-PGP). Fig. S13. Schematic illustration of preparation of PPV-loaded PEGylated LCD NPs (LAD-PEG NPs) by a nanoprecipitation/self-assembly method. Fig. S14. Ex vivo imaging of GFP fluorescence intensities in lung tissues of mice at week 3 after i.v. inoculation of 4T1-GFP tumor cells. Fig. S15. In vivo fluorescence images of mice inoculated with 4T1-GFP cells by i.v. injection. Fig. S16. Flow cytometric profiles showing neutrophil counts in bronchoalveolar lavage fluid (BALF) from mice with or without inoculation of 4T1 cells. Fig. S17. Analysis of correlation between the luminescence intensity and the H2O2 concentration in BALF. Fig. S18. Luminescence imaging of acute inflammation in the lungs with LAD-PGP NPs. Fig. S19. Comparison of peripheral blood neutrophils in mice with lung metastasis or lung infection. Fig. S20. Cytotoxicity evaluation of LAD NPs. Fig. S21. Changes in mouse body weight after a single i.v. administration of LAD NPs at 500 or 1000 mg/kg. Fig. S22. The organ index of typical major organs at day 15 after treatment with LAD NPs. Fig. S23. The blood levels of WBC, RBC, PLT, and HGB at day 15 after treatment with various doses of LAD NPs. Fig. S24. Serum levels of ALT, AST, UREA, and CREA at day 15 after treatment with various doses of LAD NPs. Fig. S25. H&E-stained pathological sections of typical major organs.