Additional file 1 of Biomimetic manganese-eumelanin nanocomposites for combined hyperthermia-immunotherapy against prostate cancer
Creators: name: Liu, Yu
name: Shang, Wenting
name: Liu, Heng
name: Hui, Hui
name: Wu, Jun
name: Zhang, Wei
name: Gao, Pengli
name: Guo, Kunxiong
name: Guo, Yanli
name: Tian, Jie
Publisher: name: figshare
Year: 2022
Subjects: subject: Immunology
subject: FOS: Clinical medicine

description: Additional file 1: Figure S1. EDX analysis of RMnMels. Figure S2. (a)XPS survey spectra. XPS spectra of (b) C1s, (c) N1s and (d) O1s for RMnMels. Figure S3. (a) Hydrodynamicsizes of nanocomposites. (b) Zeta potential of nanocomposites. Figure S4. The linear relationship for the r1relaxivity of RMnMels versus Mnconcentrations. The insets show corresponding T1-weighted MRimages. Figure S5. (a) T1-weightedMR images of mice kidneys and liver prior to and at various time points postintravenous injection of RMnMels at 12 mg/kg body weight. The images wereacquired at 1.5 T.Normalized signal intensities determined from mice (b) kidneys and (c) liver. Figure S6. UV–vis absorption profile ofRMnMels with various mass concentrations. Figure S7. Invitro photoacoustic signal intensities of RMnMels versus solution mass concentrations.The insets show corresponding photoacoustic images. Figure S8. The amount of manganese (Mn) elements per gramof tissue. Figure S9. Cell viability ofPC3 and RAW 264.7 cells after co-incubation with various concentrations ofRMnMels. Figure S10. (a) Optical microscope images of RAW 264.7cells after co-incubation with various concentrations of RMnMels for 12 h. Scale bars, 100 µm. (b) Opticalmicroscope images of PC3 cells after incubation without (upper) or with (lower)50 µg/mL RMnMels for 12 h. Scale bars, 100 µm. FigureS11. (a) Cellular TEM images ofPC3 cells after incubation without (upper) or with (lower) RMnMels at 50 µg/mLfor 12 h. (b) Tissue TEM images of PC3 tumor tissue slices after intravenous injection of PBS(upper) or RMnMels (lower) for 48 h. Figure S12. Cellular TEM imagesof lipopolysaccharide induced M1-like macrophage after incubation withRMnMels at 50 µg/mL for 12 h. Figure S13. (a) TEM images ofRMnMels with/without 2.5 mM H2O2 for 72 h. Scale bar, 200 nm.The insets show corresponding digital photographs. (b) UV–vis absorption spectra of RMnMels co-incubation with 2.5 mM H2O2 atdifferent time points. Figure S14. (a) Infrared thermalimages and (b) temperaturevariations of 200 μg/mL RMnMels aqueous solution during exposure to a 690 nm laser with different powerdensities for 300 s. (c) Infrared thermal images and (d) temperature variations of RMnMels aqueous solution with differentconcentrations during exposure to a 690 nm laser at 500 mW/cm2for 300 s. Figure S15. Photothermal heating curvesof RMnMels (100 μg/mL) irradiated by 690 nm laser at 500 mW/cm2 over three laser ON/OFF cycles. FigureS16. (a) TEM images of RMnMels solution before and after 690 nm laserirradiation (500 mW/cm2, 30 min). Scale bar, 200 nm. The insets showcorresponding digital photographs. (b) UV–vis absorption spectra of RMnMels aqueoussolutions prior to and after exposure to 690 nm laser irradiation. FigureS17. Flow cytometric assay of the polarization ofRAW 264.7 cells toward M1 phenotype. Figure S18. Body weight curves of PC3 tumor-bearing mice during a periodof 19 days after different treatments. FigureS19. Flow cytometry analysis of tumor immune cells. Gating strategy showingdelineation of (a) singlets cells. (b) numbers indicate the percentages of thecells within the gates. (c) live cells. (d) CD45+ leukocytes. (e) tumor-associatedmacrophages. Figure S20. H&amp;Estained images of mice major organ sections on day 19 after differenttreatments. Scale bar, 100 µm. FigureS21. Serum biochemical indexes of healthy mice obtained on different timepoint. Table S1. Primer sequences for different genes.