Additional file 1 of Biomimetic GBM-targeted drug delivery system boosting ferroptosis for immunotherapy of orthotopic drug-resistant GBM
Creators: name: Liu, Bao
name: Ji, Qifeng
name: Cheng, Ying
name: Liu, Miao
name: Zhang, Bangle
name: Mei, Qibing
name: Liu, Daozhou
name: Zhou, Siyuan
Publisher: name: figshare
Year: 2022
Subjects: subject: Biophysics
subject: Space Science
subject: Medicine
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Neuroscience
subject: Pharmacology
subject: Biotechnology
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Developmental Biology
subject: Cancer
subject: Hematology
subject: Infectious Diseases
subject: FOS: Health sciences
subject: Virology

description: Additional file 1: Figure S1. Diagram of the applied magnetic field in orthotopic drug-resistant GBM mice brain after Fe3O4-siPD-L1@M-BV2 was injected into the tail vein. Figure S2. Characterization of Fe3O4-siPD-L1@M-BV2. (A) Element mapping analysis diagram of Fe3O4. (B) The zeta potential of Fe3O4, Fe3O4-siPD-L1, M-BV2 and Fe3O4-siPD-L1@M-BV2. (C) Stability of Fe3O4-siPD-L1@M-BV2 in water, PBS and 10% FBS solution. (D) TEM image of Fe3O4. (E) TEM image of Fe3O4-siPD-L1@M-BV2. (F) Hemolysis phenomenon of Fe3O4-siPD-L1@M-BV2. (G) Statistic analysis of hemolysis of Fe3O4-siPD-L1@M-BV2. (n = 3, mean ± SD). Figure S3. The density of GL261/TR cells, HT-22 cells, BV2 cells and RAW264.7 cells after the cells incubated at 37 ℃ for 24 h. (A) The density of GL261/TR cells, HT-22 cells, BV2 cells and RAW264.7 cells observed by optical microscope. (B) The density of GL261/TR cells, HT-22 cells, BV2 cells and RAW264.7 cells counted by using cell counters. (n = 3, mean ± SD, ns: no significant difference). Figure S4. Statistic analysis of Fe3O4-FAM@M-BV2 uptake by GL261/TR cells, HT-22 cells, BV2 cells and RAW264.7 cells detected by flow cytometer (n = 3, mean ± SD, *P &lt; 0.05, **P &lt; 0.01, ns: no significant difference). Figure S5. The uptake mechanism of Fe3O4-FAM@M-BV2 on GL261/TR cells detected by flow cytometer. (A) Effects of different inhibitors on the uptake of Fe3O4-FAM@M-BV2 by GL261/TR cells. (B) Statistic analysis of Fe3O4-FAM@M-BV2 uptake by GL261/TR cells (n = 3, mean ± SD, **P &lt; 0.01). Figure S6. The effect of Fe3O4-siPD-L1@M-BV2 on protein expression of PD-L1 in GL261 cells. (n = 3, mean ± SD, *P &lt; 0.05, **P &lt; 0.01). Figure S7. The effect of Fe3O4-siPD-L1@M-BV2 on the viability of GL261/TR cells. (A) The effect of Fe3O4-siPD-L1@M-BV2 on the death and living GL261/TR cells. (B) Statistic analysis of death and living GL261/TR cells. (n = 3, mean ± SD, *P &lt; 0.05, **P &lt; 0.01, ns: no significant difference). Figure S8. The effect of IFN-γ on the xCT protein expression in GL261/TR cells. (n = 3, mean ± SD, **P &lt; 0.01). Figure S9. The resistance values between transwell donor chamber and recipient chamber within 12 h after drug administration. (n = 3, mean ± SD, ns: no significant difference). Figure S10. Statistic analysis of Fe3O4-FAM@M-BV2 uptake by GL261/TR cells after Fe3O4-FAM@M-BV2 penetrated in vitro BBB detected by flow cytometer. (n = 3, mean ± SD, **P &lt; 0.01). Figure S11. The distribution of Fe3O4-siPD-L1@M-BV2 in organs of orthotopic drug-resistant GBM mice observed by in vivo bioluminescence imaging. Figure S12. The effect of Fe3O4-siPD-L1@M-BV2 on body weight of orthotopic drug-resistant GBM mice. (n = 10, mean ± SD). Figure S13. The expression of invasion-related proteins in orthotopic drug-resistant GBM tissue after orthotopic drug-resistant GBM mice was treated with Fe3O4-siPD-L1@M-BV2. (A) The expression of invasion-related proteins in orthotopic drug-resistant GBM tissue detected by western blot. (B) Semi-quantitative analysis of invasion-related proteins. (n = 3, mean ± SD, *P &lt; 0.05, **P &lt; 0.01). Figure S14. Effect on the protein expression of GPX4 and PD-L1 in orthotopic drug-resistant GBM tissue after orthotopic drug-resistant GBM mice was treated with Fe3O4-siPD-L1@M-BV2. (A) Semi-quantitative analysis of GPX4 protein expression in orthotopic drug-resistant GBM tissue detected by immunofluorescence staining. (B) Semi-quantitative analysis of PD-L1 protein expression in orthotopic drug-resistant GBM tissue. (n = 3, mean ± SD, *P &lt; 0.05, **P &lt; 0.01, ns: no significant difference). Figure S15. Typical flow cytometric graph of the matured DC cells. Figure S16. Typical flow cytometric graph of CD3+CD8+IFN-γ+ T cells. Figure S17. Typical flow cytometric graph of CD4+CD25+FoxP3+ T cells.