Additional file 1 of Double-activation of mitochondrial permeability transition pore opening via calcium overload and reactive oxygen species for cancer therapy
Creators: name: Zhou, Ying
name: Jing, Shisong
name: Liu, Sainan
name: Shen, Xizhong
name: Cai, Lihan
name: Zhu, Changfeng
name: Zhao, Yicheng
name: Pang, Maolin
Publisher: name: figshare
Year: 2022
Subjects: subject: Biophysics
subject: Biochemistry
subject: Medicine
subject: Cell Biology
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Cancer

description: Additional file 1: Table S1. Size of Nanoparticles. Figure S1. The SEM images of FeCOF. Figure S2. The SEM images of FeCOF@CaCO3. Figure S3. (a) XPS spectra of FeCOF@CaCO3. (b) XPS high-resolution scans of Ca 2p. Figure S4. The PXRD of FeCOF and FeCOF@CaCO3. Figure S5. (a) N2 adsorption and desorption isotherms at 77 K of FeCOF and FeCOF@CaCO3. (b)Pore size distribution plots of FeCOF and FeCOF@CaCO3. Figure S6. The Hydrodynamic diameters of FeCOF, FeCOF@CaCO3 and FCCF. Figure S7. The zate potential of FeCOF, FeCOF@CaCO3 and FCCF. Figure S8. (a) The FCCF dispersed in water, PBS and cell culture medium (containing 10% serum) before and after 7 days incubation. (b) The TEM image of FCCF stored in cell culture medium (containing 10% serum) for a week. (c) The DLS was measured on day 0, 5 and 7 when FCCF was dispersed in water, PBS and cell culture medium (containing 10% serum). Figure S9. (a) SEM images of FCCF at pH 7.4, 6.5 and 5.5, related to Figure 2a. Data are presented as mean ± SD (n = 3). Figure S10. (a) SEM images of FeCOF at pH 7.4, 6.5 and 5.5. Data are presented as mean ± SD (n = 3). Figure S11. Oxygen release behaviour of OFCCF in solution. Figure S12. CLSM microimages of intracellular Ca2+ content in 4T1 cells. Figure S13. CLSM microimages of mitochondrial Ca2+ content in 4T1 cells. Figure S15. Quantitative analysis of the intracellular O2 generation of 4T1 cells after various treatments based on the confocal images shown in Figure 3b by using imageJ, related to Figure 3b. Figure S16. Quantitative analysis of HIF-1α protein expression, as the ratio of protein to β-actin from Western Blot results. Related to Figure 3c. P values were calculated by one-way analysis (*p&lt;0.05, **p&lt;0.01, ***p&lt;0.001). Figure S17. Intracellular ROS production of 4T1 cells under normoxic and hypoxic conditions after various treatments. Figure S18. MPTP opening in mitochondria of 4T1 cells after different treatments. Figure S19. mitochondrial membrane potential images of 4T1 cells after different treatments. Figure S20. Changes of intracellular ATP contents of 4T1 cells after being treated with PBS, L, COF+, FCCF, FCCF+ and OFCCF+ for 24 h. P values were calculated by one-way analysis (*p&lt;0.05, **p&lt;0.01, ***p&lt;0.001). Data were represented as mean ± SD (n = 4). Figure S21. Relative viabilities of L929 cells treated by incubation with COF and FCCF at different concentration for (a) 24 h and (b) 48 h. Data were represented as mean ± SD (n = 6). Figure S22. The images of 4T1 cells after various treatments and stained with calcein-AM (live cells: green) and PI (dead cell: red). Figure S23. Quantitative analysis of Annexin V-FITC/PI co-stained 4T1 cells at 24 h after various treatments. Figure S24. Changes of body weight of mice in different groups after treatment. Figure S25. The tumor picture of mice before and after treatment in different groups (n = 8). Figure S26. (a) The photos of lung at the end of various treatments. (b) H&amp;E-stained lung sections at the end of various treatments. Red arrows indicate the sites of tumor metastasis. Figure S27. Quantitative analysis of caspase 3, Bcl-2 and Cyt c protein expression, as the ratio of protein to β-actin from Western Blot results. Related to Figure 6 (f). Figure S28. ATP contents of tumor after being treated with PBS, L, COF+, FCCF, FCCF+ and OFCCF+. P values were calculated by one-way analysis (*p&lt;0.05, **p&lt;0.01, ***p&lt;0.001). Data are presented as mean ± SD (n = 4). Figure S29. (a) The HIF-1α protein expression of tumor via western blotting analysis. (b) Quantitative analysis of HIF-1α protein expression, as the ratio of protein to β-actin from Western Blot results. P values were calculated by one-way analysis (*p&lt;0.05, **p&lt;0.01, ***p&lt;0.001). Figure S30. H&amp;E stained images of the major organs, including heart, liver, spleen, lung, and kidney. Scale bar: 20 μm. Figure S31. BALB/c mice were sacrificed after PBS and FCCF (NPs) treatment. Serum biochemistry data including Alanine aminotransferase (ALT), Aspartate aminotransferase (AST) and Alkaline phosphatase (ALP) as hepatic function indicators and Blood urea nitrogen (BUN) as renal function indicators were measured. Blood hematological counts: Blood levels of White blood cells (WBC), Red blood cells (RBC), Hemoglobin (HGB), Hematocrit (HCT), Mean corpuscular volume (MCV), Mean corpuscular hemoglobin (MCH), Mean corpuscular hemoglobin concentration (MCHC) and Blood platelet (PLT). Data are presented as mean ± SD (n = 4).