2026-10-12T20:47:11Z https://api.figshare.com/v2/oai

oai:figshare.com:article/34040289 2026-10-01T06:09:50Z category_4 category_7 category_12 category_14 category_16 category_873 category_734 category_62 category_64 portal_81 item_type_6 month_year_10_2026

Additional file 1 of MHY@MO@PPP inhibits ferroptosis positive feedback and delays the acute kidney injury-to-chronic kidney disease transition by remodeling the Fe2+-mTORC1-GPX4 axis Chenhao Li (822769) Chao Zhang (51048) Chuanqi Zhang (1639483) Shiyu Duan (15310003) Jiali Zhang (31748) Hongyue Wang (2009332) Biochemistry Medicine Cell Biology Molecular Biology Physiology Chemical Sciences not elsewhere classified Biological Sciences not elsewhere classified Marine Biology Cancer Ferroptosis Acute kidney injury-to-chronic kidney disease transition Mechanistic target of rapamycin complex 1 (mTORC1) Glutathione peroxidase 4 (GPX4) Molybdenum Oxide Nanozyme Fe2+-mTORC1-GPX4 Axis Supplementary Material 1: Figure S1. Construction process and physicochemical property analysis of MoOx nanozymes and protein-environment stability of MHY@MO@PPP. Note: (A) Schematic diagram illustrating the construction process of MoOx nanozymes; (B) TEM observation of the overall morphology and dispersion state of MoOx nanozymes, scale bar: 500 nm; (C) DLS analysis of MoOx nanozymes; (D) FT-IR detection of the chemical bond structure and surface functional group characteristics of MoOx nanozymes; (E) XPS analysis of the chemical valence composition of Mo (Mo⁶⁺/Mo⁵⁺) in MoOx nanozymes, used to evaluate oxygen-vacancy structural characteristics; (F) UV-Vis-NIR absorption spectrum detection of the light absorption characteristics of MoOx nanozymes in the visible to NIR-II range; (G) continuous monitoring for 30 days at 4°C to observe the storage dispersibility of the nanozymes; (H) DLS detection of the hydrodynamic size distribution of MHY@MO@PPP after incubation in PBS or PBS+10% FBS; (I) Zeta potential detection of MHY@MO@PPP after incubation in PBS or PBS+10% FBS; (J) DLS monitoring of MHY@MO@PPP hydrodynamic diameter in PBS+10% FBS at 0, 6, 12, and 24 h to evaluate short-term colloidal stability in a protein-containing environment. All experiments were independently repeated three times, and quantitative data are presented as mean ± SD where applicable. Figure S2. Encapsulation and release characteristics of MHY1485 in MHY@MO@PPP nanozyme complex. Note: (A) LC-MS/MS detection of the characteristic peak of MHY1485 in the MHY@MO@PPP nanozyme complex to verify the encapsulation of MHY1485; (B) LC-MS/MS detection of the release behavior of MHY1485 from the MHY@MO@PPP nanozyme complex under different pH conditions; (C) LC-MS/MS detection of the release behavior of MHY1485 from the MHY@MO@PPP nanozyme complex under different H2O2 stimulation conditions; (D) Laser scanning microscopy observation of the uptake of nanomaterials by HK-2 cells; red fluorescence represents Dil, and blue is DAPI nuclear staining (scale bar: 25 μm). Figure S3. Construction and phenotypic analysis of the HK-2 cell ferroptosis model under a high iron environment. Note: (A) JC-1 fluorescence staining detected mitochondrial membrane potential changes in HK-2 cells, bar: 25 μm; (B) DCFH-DA fluorescence probe detected intracellular ROS generation levels in HK-2 cells, bar: 25 μm; (C) Biochemical kit detected intracellular MDA content in HK-2 cells; (D) Biochemical kit detected intracellular GSH content in HK-2 cells; (E) CCK-8 assay detected changes in HK-2 cell viability; (F) IF staining combined with laser confocal microscopy detected the expression and distribution of Fibronectin protein in HK-2 cells, bar: 25 μm. Cell experiments were repeated three times. * indicates comparison between two groups, p < 0.05, **p < 0.01, ***p < 0.001. Figure S4. Analysis of iron homeostasis dysregulation and cell death characteristics in the ferroptosis model. Note: (A-B) FerroOrange fluorescent staining detection of intracellular free iron levels in HK-2 cells (A) and quantitative analysis results (B), bar: 25 μm; (C) Flow cytometry Annexin V/PI staining detection of HK-2 cell death proportion; (D) Quantitative analysis of PI-positive cell distribution in HK-2 cells by flow cytometry. Cell experiments were repeated three times. * indicates comparison between two groups, p < 0.05, ***p < 0.001. Figure S5. Analysis of molecular expression profiles and renal injury markers in the ferroptosis model. Note: (A) Western blot detection of ferroptosis-related proteins (GPX4, ACSL4, FTH1) expression in HK-2 cells; (B) Western blot detection of autophagy-related proteins p-mTOR and LC3II/I expression in HK-2 cells; (C) Western blot detection of oxidative stress-related protein SOD2 expression in HK-2 cells; (D) RT-qPCR detection of mRNA expression of renal tubular injury markers KIM1, NGAL, and CYSC in HK-2 cells; (E) RT-qPCR detection of mRNA expression of fibrosis and EMT-related genes ACTA2, Fn1, and VIMENTIN in HK-2 cells. Cellular experiments were repeated three times. * indicates comparison between two groups, p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Figure S6. Effects of MHY@MO@PPP nanocomposites on iron ion homeostasis and ferroptosis in cells. Note: (A-B) FerroOrange detection of Fe2+ levels in HK-2 cells, bar: 25 μm; (C-D) Annexin V/PI flow cytometry detection of Annexin V/PI-positive cell death proportion. Cell experiments were repeated three times. Comparisons were analyzed by one-way ANOVA followed by Tukey's post hoc test. *p < 0.05, **p < 0.01, ***p < 0.001. Figure S7. Detection of cellular Fe2+ load, death-related phenotypes, and renal injury/fibrosis markers after intervention of the Fe2+-mTORC1-GPX4 axis. Note: (A-B) FerroOrange detection of Fe2+ levels in HK-2 cells, scale bar: 25 μm; (C-D) Annexin V/PI flow cytometry detection of death-related Annexin V/PI-positive cells; (E) RT-qPCR detection of renal injury markers KIM1, NGAL, and CYSC; (F) RT-qPCR detection of fibrosis/EMT markers ACTA2, Fn1, and VIMENTIN. Cell experiments were repeated three times. * indicates comparison between two groups, p < 0.05, **p < 0.01, ***p < 0.001. 2026-08-20T05:00:00Z Text Journal contribution 10.6084/m9.figshare.34040289.v1 https://figshare.com/articles/journal_contribution/Additional_file_1_of_MHY_MO_PPP_inhibits_ferroptosis_positive_feedback_and_delays_the_acute_kidney_injury-to-chronic_kidney_disease_transition_by_remodeling_the_Fe2_-mTORC1-GPX4_axis/34040289 CC BY