Additional file 1 of Iron oxide@chlorophyll clustered nanoparticles eliminate bladder cancer by photodynamic immunotherapy-initiated ferroptosis and immunostimulation
Creators: name: Chin, Yu-Cheng
name: Yang, Li-Xing
name: Hsu, Fei-Ting
name: Hsu, Che-Wei
name: Chang, Te-Wei
name: Chen, Hsi-Ying
name: Chen, Linda Yen-Chien
name: Chia, Zi Chun
name: Hung, Chun-Hua
name: Su, Wu-Chou
name: Chiu, Yi-Chun
name: Huang, Chih-Chia
name: Liao, Mei-Yi
Publisher: name: figshare
Year: 2022
Subjects: subject: Biochemistry
subject: Medicine
subject: Cell Biology
subject: Molecular Biology
subject: Pharmacology
subject: Evolutionary Biology
subject: FOS: Biological sciences
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Cancer
subject: Infectious Diseases
subject: FOS: Health sciences

description: Additional file 1:Figure S1. EDS measurement of the elemental composition ofthe Fe3O4@Chl/Cu CNPs. Figure S2. Raman spectra of the Fe3O4@Chl/FeCNPs synthesized with different Chl/Fe concentrations, capping-free Fe3O4nanocrystals, and Chl/Fe molecules. Figure S3. (a) DLS, (b) zeta potential, and (c–f) the sizedistribution of cluster particles of Fe3O4@Chl/Fe CNPssynthesized with different Chl/Fe concentrations. Figure S4. SQUID measurement of Fe3O4@Chl/Fep[0–200 mg] CNPs at 300 K. Figure S5. (a) Photographs and (b–e) TEM images of Fe3O4nanocrystal solutions with different citric acid concentrations. TEM images:(b) 2.1 mg (c) 10.6 mg (d) 21.2 mg (e) 42.4 mg. Figure S6. X-ray diffraction pattern of Fe3O4nanocrystals synthesized by using different citrate acid concentrations. Figure S7. RNO/imidazole assay for theproduction of singlet oxygen species at 25 ppm[Fe] from the Fe3O4@Chl/FeCNPs, Fe3O4, Chl/Fe, and physically mixed Fe3O4+Chl/Femolecules upon 660 nm light irradiation for 10 min. Figure S8. Thermal cycle curves of the Fe3O4@Chl/Fep[200 mg] CNPs ([Fe] = 100 ppm) under (a) 650 nm and (b) 808 nm laserirradiation. (c) Thermal cycle curves of Chl/Fe ([Chl/Fe] = 0.2 mM) under 650 nm and 808 nm laser irradiation. Figure S9. In vitro characterizations of the chromogenic performance of a varietyof Fe3O4@Chl/Fe[0–200 mg] CNPs ([Fe] = 0.5 ppmfrom AAS and [Chl/Cu] = 0.001 mM from UV–Visible measurements). Fe3O4–N2H4is the iron oxide from reference 45. (b) TMB assay for the reaction of iron-basedmaterials at 0.5 ppm[Fe] with 1 mM H2O2. (c) UV–Vis spectra of Fe3O4@Chl/Fe[200 mg] CNPs at 0.5 ppm[Fe]. Figure S10. TMB assay for evaluating the reaction of H2O2with (a) 0.5 ppm[Fe] Fe3O4@Chl/Fe[200 mg]CNPs and 0.5 ppm[Fe] ppm[Fe] Fe3O4@Chl/Fe[200 mg] CNPs plus 1 mM GSH and (b) 0.5 ppm[Fe] Chl/Fe molecule andChl/Fe molecule plus 1 mM GSH. Figure S11. GSH depletion assay under (a) 20 ppm[Fe]Fe-based treatments and (b) 20 ppm[Fe] Fe-based treatments combinedwith a 100 μM H2O2 solution. Figure S12. GSH depletion assay with 20 ppm Fe3O4@Chl/Fe[200 mg] CNPs and Fe3O4@Chl/Fe[200 mg]-CPBACNPs combined with irradiation by a 75 mW/cm2 660 nm laser. Figure S13. AAS measurements for theuptake analysis of (a) T24 cancer cells, (b) MB49 cancer cells, (c) SV-HUC1normal cells, and (d) Vero normal cells after treatment with Fe3O4@Chl/Fe[200 mg] and Fe3O4@Chl/Fe[200 mg]-CPBA CNPsfor 1, 16 and 24 h. Figure S14.Confocal images of T24 cells treated with Fe3O4@Chl/Fe[200 mg]-CPBA CNPs. (a) Fluorescence and confocal images from bottom to top (b–d).(scale bar: 20 μm). Figure S15. Cellviability after treated with different concentrations of modified Fe3O4@Chl/Fe[200 mg] CNPs with targeting molecules such as CPBA, FA, RGD, and transferrin(a) without and (b) with a 660 nm LED light source are displayed. Figure S16. The cell viability of HeLa,NIH 3T3 and VERO cells treated with Fe3O4@Chl/Fe[200 mg] CNPs group and CPBA-, RGD-, andtransferrin-conjugated Fe3O4­@Chl/Fe[200 mg] CNPs without (a, c, e) and with (b, d, f) a 660 nmLED light source at 75 mW/cm2. Figure S17. The cell viability of MB49 treated with Fe3O4@Chl/FeCNPs and Fe3O4@Chl/Fe-CPBA CNPs (a) without and (b) withirradiation by a 660 nm LED light source at 75 mW/cm2. Figure S18.The viability of T24 cells treated with Fe3O4@Chl/Fe[5 mg and 200 mg] CNPs synthesized with 5 and 200 mg of Chl/Fe for 24 h. (a)Without light exposure (b) exposed to 660 nm LED light (75 mW/cm2)([Fe] = 0.2, 1, 5, 10, 50, 100 ppm). Figure S19. Confocal images of MB49 cells treated with Fe3O4@Chl/Fe[200 mg]-CPBA CNPs after24 h: (a) Fluorescence and confocal images from bottom to top (b–d). Scale bar:20 μm.