Additional file 1 of A multifunctional AIE gold cluster-based theranostic system: tumor-targeted imaging and Fenton reaction-assisted enhanced radiotherapy

Additional file 1: Fig. S1. PXRD patterns of Au4 cluster. Fig. S2. TEM image of oil-phase IO NCs. Fig. S3. DLS spectra of (a) PEG modified IO NC and Au4 cluster in DMF with (b) 0% PBS, (c) 20% PBS, (d) 40% PBS, and (e) 60% PBS, (f) 80% PBS. Fig. S4. DLS spectra of (a) Au4-IO NP in DMF/ethanol with 60% medium and (b) 1000 times diluted Au4-IO NP using cell medium. Fig. S5. Emission spectra and (b) DLS spectra of Au4-IO NP-cRGD in serum at day 0, 3, 5, and 7. Fig. S6. Zeta potential of Au4-IO NP and Au4-IO NP-cRGD. Fig. S7. 1H-NMR spectra of cRGD-PEG-COOH. The characteristic peak of cRGD and PEG confirm the structure of cRGD-PEG-COOH. Fig. S8. 1H-NMR spectrum and DOSY spectrum of cRGD-PEG-COOH. Fig. S9. Flow cytometric assay of 4T1 cells after incubation with PBS, Au4-IO NP and Au4-IO NP-RGD. Fig. S10. In vitro cytotoxicity against 4T1 cells of Au4-IO NP and Au4-IO NP-cRGD with 4 Gy X-ray irradiation. Fig. S11. Confocal imaging of 4T1 cells after incubation with Au4 and IO NC. Scale bar: 50 μm. Fig. S12. Viability of 4T1 cells in different treatment groups: (1) Au4-IO NP-cRGD, (2) pH 6.5, (3) H2O2, (4) Au4-IO NP-cRGD pH 6.5, (5) Au4-IO NP-cRGD + H2O2, and (6) Au4-IO NP-cRGD + H2O2 pH 6.5. Fig. S13. Live/dead imaging of 4T1 cells after receiving different treatments (2 Gy X-ray, Au4-IO NP + 2 Gy X-ray, and Au4-IO NP-cRGD + 2 Gy X-ray). Green: live; red: dead. Scale bar: 50 μm. Fig. S14. Flow cytometric assay of 4T1 cells (negative control, positive control). Fig. S15. Flow cytometric assay of 4T1 cells with different treatments (batch 1). Fig. S16. Flow cytometric assay of 4T1 cells with different treatments (batch 2). Fig. S17. Flow cytometric assay of 4T1 cells with different treatments (batch 3). Fig. S18. Representative images of the colony formation assay of 4T1 cells with different treatments. Fig. S19. Survival curve of 4T1 cells received Au4-IO NP. Fig. S20. The fluorescence intensity of hydroxyl radical imaging of 4T1 cells (λex = 540 nm) at 6 h after administering different treatments. (1) Control, (2) Au4-IO NP-cRGD, (3) 4 Gy, (4) Au4-IO NP + 4 Gy, and (5) Au4-IO NP-cRGD + 4 Gy. Fig. S21. Confocal images of γ-H2AX expression in 4T1 cells after receiving various treatments. Scale bar: 50 μm. Fig. S22. The fluorescence intensity of γ-H2AX imaging of 4T1 cells (λex = 647 nm) at 6 h after administering different treatments. (1) Control, (2) Au4-IO NP-cRGD, (3) 4 Gy, (4) Au4-IO NP + 4 Gy, and (5) Au4-IO NP-cRGD + 4 Gy. Fig. S23. The fluorescence intensity of Caspase-3/7 imaging of 4T1 cells (λex = 488 nm) at 6 h after administering different treatments. (1) Control, (2) Au4-IO NP-cRGD, (3) 4 Gy, (4) Au4-IO NP + 4 Gy, and (5) Au4-IO NP-cRGD + 4 Gy. Fig. S24. The biodistribution of (a) Au4-IO NP and (b) Au4-IO NP-cRGD at 6 h, 24 h, 48 h, 72 h, and 7 d after intravenous injection. Fig. S25. (a) Images of dissected tumors in control, Au4-IO NP-cRGD, and Au4-IO NP-cRGD + 4 Gy. (b) Images of dissected tumors in 4 Gy group. Fig. S26. (a) Representative images of mice of Au4-IO NP + 4 Gy group at days 0 and 14. (b) Tumor volume curve of the mice. (c) Mouse body growth curve. (d) Image of dissected tumors in Au4-IO + 4 Gy.