DOI: 10.5061/dryad.rn8pk0pn4
title: Immuno-engineered mitochondria for efficient therapy of acute organ injuries via modulation of inflammation and cell repair

description: Acute organ injuries are a major public health concern, driven by
 inflammation and mitochondrial dysfunction, leading to cell damage and
 organ failure. In this study, we engineered neutrophil membrane-fused
 mitochondria (nMITO), which combine the injury-targeting and
 anti-inflammatory properties of neutrophil membrane proteins with the
 cell-repairing function of mitochondria. nMITO effectively blocked
 inflammatory cascades and restored mitochondrial function, targeting both
 key mechanisms in acute organ injuries. Additionally, nMITO selectively
 targeted damaged endothelial cells via β-integrins and was delivered to
 injured tissues through tunneling nanotubes, enhancing their regulatory
 effects on inflammation and cell damage. In mouse models of acute
 myocardial injury, liver injury, and pancreatitis, nMITO significantly
 reduced immune dysfunction and repaired tissue damage. These findings
 suggest that nMITO is a promising therapeutic strategy for managing acute
 organ injuries.
description: # Immuno-engineered mitochondria for efficient therapy of acute organ
 injuries via modulation of inflammation and cell repair
 [https://doi.org/10.5061/dryad.rn8pk0pn4](https://doi.org/10.5061/dryad.rn8pk0pn4) This dataset comprises the processed data tables corresponding to the main experimental figures and supplementary materials presented in the Science Advances article “Immuno-Engineered Mitochondria for Efficient Therapy of Acute Organ Injuries via Modulation of Inflammation and Cell Repair.” The data include quantitative measurements, imaging analyses, and in vivo/in vitro experimental results that evaluate mitochondrial engineering, inflammation modulation, and cell repair processes. ## Description of the data and file structure The **data.zip** archive contains the following Excel files, each representing processed data for specific figures in the publication: ### **1.    Figure 1 Preparation and Characterization.xlsx** **Preparation and Characterization of Neutrophil Membrane Fused Mitochondria (nMITO)** This file contains data related to the preparation and comprehensive characterization of nMITO. The experimental design and schematic illustrations are detailed in the article. The figure comprises the following panels: **Fig1C. Dynamic Light Scattering (DLS) Measurements** * **Units:** Nanometers (nm) / Zeta potential (mV) * **Description:** Measurements of the hydrodynamic size (diameter) and the zeta potential of MITO, NEM, and nMITO using DLS. **Fig1F. Mitochondrial Membrane Potential (MMP) Measurement** * **Units:** Arbitrary units (a.u.) * **Description:** Assessment of the mitochondrial membrane potential of nMITO using Tetramethylrhodamine, methyl ester (TMRM) fluorescence. **Fig1G. ATP Synthesis Activity** * **Units:** Relative units (e.g., relative ATP production) * **Description:** Measurement of ATP synthesis activity to evaluate the energy metabolism of mitochondria. **Fig1H. Oxygen Consumption** * **Units:** nmol/min/mg protein * **Description:** Measurement of the oxygen consumption rate of mitochondria, reflecting their respiratory activity. **Fig1J–K. Neutralization Curves** * **Units:** pg/mL * **Description:** Neutralization curves demonstrating the ability of nMITO to neutralize inflammatory cytokines (Panel J) and chemokines (Panel K). *Each sheet is organized with treatment groups as column headers (e.g., Control, Saline, MITO, nMITO) and the corresponding measured values listed in subsequent rows.* --- ### **2.    Figure 2 Targeting Ability.xlsx** **The Targeting Ability of nMITO Inherited from Neutrophil Membrane** This file contains data assessing the targeting ability of nMITO. The Excel file includes quantitative data for the following panels: **Fig2B: Uptake of nMITO and MITO by Activated HUVECs (Flow Cytometry)** * **Units:** Mean Fluorescence Intensity (MFI) * **Description:** Measured uptake of nMITO and MITO by activated HUVECs using flow cytometry. **Fig2F: Delivery of Ingested Mitochondria from HUVECs to L02 Cells (Flow Cytometry)** * **Units:** Mean Fluorescence Intensity (MFI) * **Description:** Quantitative data showing the delivery efficiency of ingested mitochondria from HUVECs to L02 cells, as monitored by flow cytometry. **Fig2H: Quantitative Fluorescent Intensity in Injured Organs** * **Units:** Mean Fluorescence Intensity (MFI) * **Description:** Measured fluorescent intensity in injured heart, liver, and pancreas tissues from mice with ISO-induced cardiac injury, APAP-induced liver injury, and CER-induced acute pancreatitis after injection of MitoTracker® Deep Red-labeled MITO, nMITO, and rMITO at 1 mg/kg. Each sheet is organized with treatment groups as column headers (e.g., Control, Saline, MITO, nMITO, rMITO) and the corresponding measured values listed in subsequent rows. --- ### **3.    Figure 3 In vitro Inflammation Neutralizing Effect.xlsx** **In vitro Inflammation Neutralizing Effect** This file contains data assessing the inflammation-neutralizing effects of nMITO in vitro using LPS-stimulated RAW264.7 macrophages and subsequent assays. The experimental design is detailed in the article. The Excel file comprises the following sheets: **Fig3B: Data for IL-1β Quantification** * **Units:** pg/mL * **Description:** Measured levels of IL-1β in the culture medium after treatment with Control, Saline, MITO, or nMITO. **Fig3C: Data for TNF-α Quantification** * **Units:** pg/mL * **Description:** Measured levels of TNF-α in the culture medium after treatment with Control, Saline, MITO, or nMITO. **Fig3D: Data for IL-6 Quantification** * **Units:** pg/mL * **Description:** Measured levels of IL-6 in the culture medium after treatment with Control, Saline, MITO, or nMITO. **Fig3E: CCK-8 Assay Results for H9c2 Cardiomyocytes** * **Units:** Percentage (%) * **Description:** Cell viability of H9c2 cardiomyocytes after exposure to supernatants from the respective treatments (Control, Saline, MITO, nMITO). **Fig3F: CCK-8 Assay Results for L02 Hepatocytes** * **Units:** Percentage (%) * **Description:** Cell viability of L02 hepatocytes following treatment with supernatants from the respective groups (Control, Saline, MITO, nMITO). **Fig3I: Quantitative Data on Neutrophil Migration in the Transwell Assay** * **Units:** Cell Count * **Description:** Number of neutrophils that migrated in the transwell assay, comparing conditions (e.g., with or without CXCL2) across treatment groups (Control, Saline, MITO, nMITO). **Fig3K: Quantitative Data on RAW264.7 Macrophage Migration in the Transwell Assay** * **Units:** Cell Count * **Description:** Number of RAW264.7 macrophages that migrated in the transwell assay under different treatments (Control, Saline, MITO, nMITO). *Each sheet is organized with treatment groups as column headers (e.g., Control, Saline, MITO, nMITO) and the corresponding measured values listed in subsequent rows. Data are presented as mean ± SD, with statistical significance indicated above.* --- ### **4. Figure 4 In Vitro Cell Repairing Activity.xlsx** **In Vitro Cell Repairing Activity of nMITO against Various Injured Cells** This file contains data evaluating the in vitro cell-repairing activity of nMITO on various injured cells. The experimental design is detailed in the article. The Excel file comprises the following sheets: **H9c2 Myocardial Cells:** * **Fig4C: Calculation of MMP in H9c2 Cells** * **Units:** Ratio (JC-1 Red/Green fluorescence intensity) * **Description:** Calculation of mitochondrial membrane potential (MMP) in H9c2 cells treated with Saline, MITO, or nMITO. * **Fig4D: ATP Levels in H9c2 Cells** * **Units:** nmol/mg protein * **Description:** Measured ATP content in H9c2 cells after treatment with Saline, MITO, or nMITO. * **Fig4E: Quantification of Myocardial Cell Hypertrophy in H9c2 Cells** * **Units:** Area (μm²) * **Description:** Quantitative analysis of myocardial cell hypertrophy in H9c2 cells based on WGA staining following treatment with Saline, MITO, or nMITO. **L02 Hepatocytes:** * **Fig4H: Calculation of MMP in L02 Cells** * **Units:** Ratio (JC-1 Red/Green fluorescence intensity) * **Description:** Calculation of mitochondrial membrane potential (MMP) in APAP-injured L02 cells treated with Saline, MITO, or nMITO. * **Fig4I: ATP Levels in L02 Hepatocytes** * **Units:** nmol/mg protein * **Description:** Measured ATP content in APAP-injured L02 hepatocytes following treatment with Saline, MITO, or nMITO. * **Fig4J: ROS Levels in L02 Hepatocytes** * **Units:** Relative expression * **Description:** Quantification of reactive oxygen species (ROS) levels in APAP-injured L02 hepatocytes after treatment with Saline, MITO, or nMITO. * **Fig4K: ALT Levels in L02 Hepatocytes** * **Units:** U/L * **Description:** Measured alanine aminotransferase (ALT) levels in APAP-injured L02 hepatocytes treated with Saline, MITO, or nMITO. * **Fig4L: AST Levels in L02 Hepatocytes** * **Units:** U/L * **Description:** Measured aspartate aminotransferase (AST) levels in APAP-injured L02 hepatocytes following treatment with Saline, MITO, or nMITO. * **Fig4N: Quantitative Data on Apoptosis in L02 Cells** * **Units:** Percentage (%) * **Description:** Flow cytometry results quantifying early and advanced apoptosis in L02 cells treated with Saline, MITO, or nMITO. **Primary Acinar Cells:** * **Fig4Q: Calculation of MMP in Primary Acinar Cells** * **Units:** Ratio (JC-1 Red/Green fluorescence intensity) * **Description:** Calculation of mitochondrial membrane potential (MMP) in primary acinar cells injured by CER or NaT and treated with Saline or nMITO. * **Fig4R: Calculation of Necrosis Ratio in Primary Acinar Cells** * **Units:** Percentage (%) * **Description:** Quantification of the necrosis ratio in primary acinar cells injured by CER or NaT following treatment with Saline or nMITO. *Each sheet is organized with treatment groups as column headers (e.g., Control, Saline, MITO, nMITO) and the corresponding measured values in subsequent rows.* --- ### **5. Figure 5 Heart in vivo.xlsx** **Therapeutic Effect of nMITO in ISO-Mediated Myocardial Injury Animal Model** This file contains data assessing the therapeutic effect of nMITO in a mouse model of ISO-mediated myocardial injury. The experimental design is detailed in the article. The Excel file comprises the following sheets: **Fig5_C: Data for HW/BW Ratio** * **Units:** Ratio * **Description:** Improvement in the heart weight to body weight (HW/BW) ratio in mice following treatment. **Fig5_D: Data for HW/TL Ratio** * **Units:** Ratio (mg/cm) * **Description:** Improvement in the heart weight to tibia length (HW/TL) ratio in mice following treatment. **Fig5_E: Data for Mitochondrial Membrane Potential** * **Units:** Mean Fluorescence Intensity (MFI) or Ratio (JC-1 Red/Green) * **Description:** Effect of different preparations (Saline, MITO, nMITO) on the mitochondrial membrane potential of mouse cardiomyocytes. **Fig5_F: Data for ATP Synthesis** * **Units:** nmol/mg protein * **Description:** ATP synthesis levels in mouse cardiomyocytes following treatment. **Fig5_G: Data for ANP mRNA Expression** * **Units:** Relative Expression (fold change) * **Description:** mRNA expression levels of ANP in heart tissue of mice after treatment. **Fig5_H: Data for BNP mRNA Expression** * **Units:** Relative Expression (fold change) * **Description:** mRNA expression levels of BNP in heart tissue of mice after treatment. *Each sheet is organized with treatment groups as column headers (e.g., Healthy, ISO, ISO + MITO, ISO + nMITO) and the corresponding measured values in subsequent rows.* --- ### **6. Figure 6 liver in vivo.xlsx** **Evaluation of nMITO on the Therapeutic Effect of APAP-Mediated Acute Liver Injury** This file contains data assessing the therapeutic effect of nMITO in a mouse model of APAP-induced acute liver injury. The experimental design is detailed in the article. The Excel file comprises the following sheets: **Fig.6B: Data for ATP Levels in Hepatocytes** * **Units:** nmol/mg protein * **Description:** Measured ATP content in hepatocytes after treatment with different preparations. **Fig.6C: Data for ROS Levels in Hepatocytes** * **Units:** Relative expression * **Description:** Measured reactive oxygen species (ROS) levels in hepatocytes following treatment with different preparations. **Fig6.D: ELISA Data for Inflammatory Factor 1** * **Units:** pg/mL * **Description:** ELISA test results for one inflammatory factor in liver tissue after treatment with different preparations. **Fig6.E: ELISA Data for Inflammatory Factor 2** * **Units:** pg/mL * **Description:** ELISA test results for a second inflammatory factor in liver tissue after treatment with different preparations. **Fig.6f: ELISA Data for Inflammatory Factor 3** * **Units:** pg/mL * **Description:** ELISA test results for a third inflammatory factor in liver tissue after treatment with different preparations. **Fig.6I: Data for ALT Levels** * **Units:** U/L * **Description:** Serum ALT levels in mice after treatment with different preparations. **Fig.6J: Data for AST Levels** * **Units:** U/L * **Description:** Serum AST levels in mice after treatment with different preparations. *Each sheet is organized with treatment groups as column headers (e.g., Control, Saline, MITO, nMITO) and the corresponding measured values in subsequent rows. * (P &lt; 0.05), ** (P &lt; 0.01), *** (P &lt; 0.001), **** (P &lt; 0.0001).* --- ### **7. Figure 7 Pancreatitis.xlsx** **Therapeutic Effect of nMITO in CER-Mediated Acute Pancreatitis Mouse Model** This file contains data assessing the therapeutic effect of nMITO in a mouse model of cerulein (CER)-mediated acute pancreatitis. The experimental design is detailed in the article. The Excel file comprises the following sheets: **Fig7C: Data for Total Pathological Scores** * **Units:** Scoring Units (arbitrary units) * **Description:** Total pathological scores of mouse pancreas after treatment with different mitochondrial preparations, based on H&E sections, F4/80 staining, and TEM images. **Fig7D: Data for Serum Lipase Concentrations** * **Units:** U/L * **Description:** Measured serum lipase levels in mice following treatment with different mitochondrial preparations. **Fig7E: Data for Serum Amylase Concentrations** * **Units:** U/L * **Description:** Measured serum amylase levels in mice following treatment with different mitochondrial preparations. **Fig7F: Data for Pancreatic Trypsin Activity** * **Units:** Relative expression * **Description:** Pancreatic trypsin activity in mice after treatment with different mitochondrial preparations. **Fig7G: Data for Serum MCP-1 Concentrations** * **Units:** pg/mL * **Description:** Serum concentrations of MCP-1 in mice following treatment with different mitochondrial preparations. **Fig7H: Data for MPO Activities in Pancreas** * **Units:** Relative expression * **Description:** MPO (myeloperoxidase) activities in mouse pancreas after treatment with different mitochondrial preparations. **Fig7I: Data for Serum TNF-α Concentrations** * **Units:** pg/mL * **Description:** Serum concentrations of TNF-α in mice following treatment with different mitochondrial preparations. *Each sheet is organized with treatment groups as column headers (e.g., Control, Saline, MITO, nMITO) and the corresponding measured values in subsequent rows. Data are presented as mean ± SD, with statistical significance indicated above.* --- ### **8. Figure 8 Severe Pancreatitis.xlsx** **Therapeutic Effect of nMITO in Severe Acute Pancreatitis Mouse Models** This file contains data assessing the therapeutic effect of nMITO in severe acute pancreatitis mouse models induced by NaT. The experimental design is detailed in the article. The Excel file comprises the following sheets: **Fig8C: Data for Pathological Scores of Mouse Pancreas** * **Units:** Scoring Units (arbitrary units) * **Description:** Pathological scores of mouse pancreas after treatment with different mitochondrial preparations. **Fig8D: Data for MPO Activities in Mouse Pancreas** * **Units:** Relative expression * **Description:** MPO (myeloperoxidase) activities in mouse pancreas following treatment with different mitochondrial preparations. **Fig8E: Data for Serum Amylase Concentrations** * **Units:** U/L * **Description:** Serum amylase levels in mice after treatment with different mitochondrial preparations. **Fig8F: Data for Serum Lipase Concentrations** * **Units:** U/L * **Description:** Serum lipase levels in mice after treatment with different mitochondrial preparations. **Fig8G: Data for Serum TNF-α Concentrations** * **Units:** pg/mL * **Description:** Serum TNF-α concentrations in mice following treatment with different mitochondrial preparations. **Fig8H: Data for Serum MCP-1 Concentrations** * **Units:** pg/mL * **Description:** Serum MCP-1 concentrations in mice after treatment with different mitochondrial preparations. **Fig8I: Data for MPO Activities in Mouse Lung** * **Units:** Relative expression * **Description:** MPO activities in the lung of mice after treatment with different mitochondrial preparations. *Each sheet is organized with treatment groups as column headers (e.g., Control, Saline, MITO, nMITO) and the corresponding measured values in subsequent rows. Data are presented as mean ± SD, with statistical significance indicated above.* --- ### **9. Supplementary Figures** This file contains additional datasets supporting the main findings of the study. The Excel file comprises the following sheets: **Fig.S1: Characterization of Isolated Mitochondria** * **Units:** * S1b: Relative Fluorescence Intensity (MFI) for TMRM * S1c: Relative ATP production (arbitrary units) * **Description:** * S1b: Quantitative data for mitochondrial membrane potential (MMP) measured via TMRM fluorescence (n = 4–5). * S1c: ATP synthesis activity of isolated mitochondria. **Fig.S2: Characterization of Neutrophils and Isolated Neutrophil Membrane (NEM)** * **Units:** Percentage (%) (for flow cytometry) * **Description:** * S2a: Flow cytometric analysis of CD11b⁺Ly6G⁺ expression in bone marrow cells and isolated neutrophils. * S2b: Data on neutrophil viability determined by PI staining (percentage of PI-positive cells; positive control with Triton X-100, negative control as NC).*(Imaging and protein extraction data are described in the manuscript.)* **Fig.S5: Characterization of nMITO Modification with Different MITO:NEM Ratios** * **Units:** * S5a: Mean Fluorescence Intensity (MFI) from FACS analysis * S5b: Encapsulation efficiency (% residual NEM) * S5c: MFI for Mitotracker Green * S5d: Percentage (%) of Mitotracker Green-negative mitochondria * S5e: ATP production (nmol/g) * **Description:** Data comparing different MITO:NEM ratios (1:1, 1:2, 1:4) regarding Ly6G expression, encapsulation efficiency, Mitotracker Green fluorescence, and ATP synthesis ability. **Fig.S6: ATP Content in MITO and nMITO Over Time** * **Units:** ATP levels (nM) * **Description:** Time-course data showing ATP content in MITO and nMITO at various time points post-preparation. **Fig.S7A: Heat Map of Membrane-Associated Proteins (Part A)** * **Units:** Relative expression (color scale; qualitative) * **Description:** Heat map illustrating relative expression levels of membrane-associated proteins in NEM and nMITO. **Fig.S7B: Heat Map of Mitochondria-Associated Proteins (Part B)** * **Units:** Relative expression (color scale; qualitative) * **Description:** Heat map illustrating relative expression levels of mitochondria-associated proteins in NEM and nMITO. **Fig.S7C: Correlation Matrix for Plasma Membrane Proteins** * **Units:** Correlation coefficient (r-value) * **Description:** Correlation matrix showing similarity in expression profiles of plasma membrane proteins between NEM and nMITO. **Fig.S7D: Quantification of Average Protein Response for Membrane-Associated Proteins** * **Units:** Arbitrary units (quantitative value) * **Description:** Quantitative analysis of average protein response values for membrane-associated proteins in NEM and nMITO. **Fig.S8A: Neutralization Curve of MITO on Proinflammatory Cytokines** * **Units:** pg/mL * **Description:** Neutralization curve showing the effect of MITO on TNF-α and CCL2 levels. **Fig.S8B: Comparison of Cytokine Neutralization Capabilities** * **Units:** pg/mL * **Description:** Comparison of the cytokine neutralization effects of MITO versus nMITO at an MITO concentration of 1 mg/mL. **Fig.S10B: Quantitative Analysis of HUVEC Uptake of nMITO** * **Units:** Percentage (%) * **Description:** Flow cytometry data showing the percentage of HUVECs that absorb MitoTracker® Green-labeled nMITO at different concentrations. **Fig.S15B: Quantitative Fluorescence Data in ISO-Induced Cardiac Injury** * **Units:** Relative Fluorescence Intensity (arbitrary units) * **Description:** Quantitative results of ex vivo fluorescence in main organs from ISO-induced cardiac injury models. **Fig.S16B: Quantitative Fluorescence Data in APAP-Induced Liver Injury** * **Units:** Relative Fluorescence Intensity (arbitrary units) * **Description:** Quantitative results of ex vivo fluorescence in main organs from APAP-induced liver injury models. **Fig.S17B: Quantitative Fluorescence Data in CER-Induced Acute Pancreatitis** * **Units:** Relative Fluorescence Intensity (arbitrary units) * **Description:** Quantitative results of ex vivo fluorescence in main organs from CER-induced acute pancreatitis models. **Fig.S19B: Co-Location Analysis of Mitochondria and ICAM-1/CD31 in Myocardium** * **Units:** Quantitative co-localization percent (%) * **Description:** Data representing the co-location analysis of mitochondria with ICAM-1 in myocardial tissue from ISO-induced cardiac injury. **Fig.S21B: Co-Location Analysis of Mitochondria and ICAM-1/CD31 in Liver** * **Units:** Quantitative co-localization percent (%) * **Description:** Data representing the co-location analysis of mitochondria with ICAM-1 in liver tissue from APAP-induced liver injury. **Fig.S23B: Co-Location Analysis of Mitochondria and ICAM-1/CD31 in Pancreas** * **Units:** Quantitative co-localization percent (%) * **Description:** Data representing the co-location analysis of mitochondria with ICAM-1 in pancreatic tissue from CER-induced acute pancreatitis. **Fig.S24: Construction of ISO-Mediated H9c2 Cell Injury Model** * **Fig.S24A: Cell Viability** * **Units:** Percentage (%) * **Description:** Viability of H9c2 cells under different concentrations of isoproterenol (ISO). * **Fig.S24B: ATP Synthesis Level** * **Units:** nmol/mg protein * **Description:** Comparison of ATP synthesis between healthy (control) and ISO-injured H9c2 cells. **Fig.S25: In Vitro Cell Repairing Activity Against Injured H9c2 Cells by nMITO** * **Fig.S25B: Mitochondrial Membrane Potential (MMP)** * **Units:** Ratio (JC-1 Red/Green) * **Description:** Calculated MMP in H9c2 cells following treatment with different concentrations of nMITO or MITO. * **Fig.S25C: ATP Synthesis Levels** * **Units:** nmol/mg protein * **Description:** ATP production in H9c2 cells after treatment with varying concentrations of nMITO or MITO. **Fig.S26: Construction of APAP-Mediated L02 Hepatocyte Injury Model** * **Fig.S26A: Cell Viability** * **Units:** Percentage (%) * **Description:** Viability of L02 hepatocytes under different concentrations of APAP. * **Fig.S26B: ALT Levels** * **Units:** U/L * **Description:** ALT enzyme levels in L02 cells under APAP treatment. * **Fig.S26C: AST Levels** * **Units:** U/L * **Description:** AST enzyme levels in L02 cells under APAP treatment. * **Fig.S26D: ATP Levels** * **Units:** nmol/mg protein * **Description:** ATP synthesis in APAP-injured L02 cells at various time points. * **Fig.S26E: ROS Levels** * **Units:** Relative expression * **Description:** ROS production in APAP-injured L02 cells measured over time. **Fig.S27: In Vitro Cell Repairing Activity Against Injured L02 Cells by nMITO** * **Fig.S27A–D:** * **Units:** * ALT/AST: U/L * ATP: nmol/mg protein * ROS: Relative expression * **Description:** Measurements of ALT, AST, ATP synthesis, and ROS levels in APAP-injured L02 cells treated with different concentrations of MITO or nMITO. * **Fig.S27F: Quantitative Analysis of MMP** * **Units:** Ratio (Red/Green) * **Description:** Quantitative evaluation of the mitochondrial membrane potential in L02 cells following treatment. **Fig.S28: Evaluation of nMITO on CER-Induced Pancreatic Acinar Cell Injury** * **Units:** * MMP: Ratio (JC-1 Red/Green) * MPTP Opening: Percentage (%) * ROS & Ca²⁺ Intensity: Relative Fluorescence Intensity (MFI) * Necrosis: Percentage (%) * **Description:** Determination of MMP, MPTP opening, ROS levels, relative Ca²⁺ intensity, and necrotic rate in CER-treated pancreatic acinar cells after Saline or nMITO treatment.*(n = 5 for JC-1/calcein-AM assays; n = 4 for ROS, Ca²⁺, and necrosis; Scale bar = 100 μm.)* **Fig.S29: Evaluation of nMITO on NaT-Induced Pancreatic Acinar Cell Injury** * **Units:** Same as Fig.S28 * **Description:** Quantitative analysis of mitochondrial function, ROS, relative Ca²⁺ intensity, and necrotic rate in NaT-treated pancreatic acinar cells after Saline or nMITO treatment.*(n = 5 for JC-1/calcein-AM assays; n = 4 for ROS, Ca²⁺, and necrosis; Scale bar = 100 μm.)* **Fig.S30: Therapeutic Effect of Different Concentrations of nMITO in ISO-Mediated Cardiac Injury** * **Fig.S30A: ATP Synthesis in Cardiomyocytes** * **Units:** nmol/mg protein * **Description:** ATP levels in cardiomyocytes from ISO-injured mice treated with varying doses of nMITO. * **Fig.S30B: Mitochondrial Membrane Potential (MMP)** * **Units:** Ratio (JC-1 aggregates/monomers) * **Description:** Dose-dependent changes in MMP in ISO-injured mice. * **Fig.S30E–F: Cardiac Function Parameters** * **Units:** * Ejection Fraction (%), Fractional Shortening (%) * **Description:** Echocardiography results in ISO-injured mice treated with different nMITO doses. * **Fig.S30G–I: Cardiac Morphometric Indices** * **Units:** Ratios (e.g., HW/BW, HW/TL, LVMI) * **Description:** Improvements in heart weight/body weight, heart weight/tibia length, and left ventricular mass index. * **Fig.S30J–N: Cardiac mRNA Expression Levels** * **Units:** Fold change (relative expression) * **Description:** Expression levels of ANP, BNP, β-MHC, type I collagen, and type III collagen in heart tissue following various nMITO doses. **Fig.S32A: Mitochondrial Membrane Potential in Mouse Cardiomyocytes** * **Units:** Ratio (JC-1 Red/Green) * **Description:** Quantitative assessment of MMP in cardiomyocytes from heart tissue of treated mice. **Fig.S33: Therapeutic Efficacy of a Single nMITO Administration in ISO-Induced Cardiac Injury** * **Fig.S33C: Cardiac Morphometric Ratios** * **Units:** Ratios (e.g., HW/BW, HW/TL) * **Description:** Quantification of cardiac hypertrophy parameters (n = 10). * **Fig.S33E: Quantitative MMP Analysis** * **Units:** Ratio (JC-1 Red/Green) * **Description:** Quantification of mitochondrial membrane potential in heart tissue. * **Fig.S33F: Cardiac Tissue ATP Levels** * **Units:** nmol/mg protein * **Description:** ATP synthesis in cardiac tissue. * **Fig.S33G–J: Echocardiography Parameters** * **Units:** Various (e.g., LVEF in %, FS in %, cardiac output, stroke volume) * **Description:** Echocardiography data showing improved cardiac function. * **Fig.S33K: Representative Echocardiographic Images** * **Description:** Images illustrating improved heart function in nMITO-treated mice. **Fig.S34: Construction of APAP-Mediated Acute Liver Injury** * **Fig.S34A–B: ALT and AST Levels** * **Units:** U/L * **Description:** Liver enzyme levels in APAP-induced liver injury. * **Fig.S34C: ROS Activities in Hepatocytes** * **Units:** Relative expression * **Description:** ROS levels in injured hepatocytes. **Fig.S35: Pancreatic Histopathology Scores (CER-AP Model)** * **Units:** Scoring Units (arbitrary) * **Description:** Histological scores reflecting pancreatic edema, inflammatory cell infiltration, and acinar cell necrosis in the CER-AP model. **Fig.S37: Pancreatic Histopathology Scores (NaT-AP Model)** * **Units:** Scoring Units (arbitrary) * **Description:** Histological scores for pancreatic tissue in the NaT-AP model. --- Each file has been formatted to clearly present the final analyzed results corresponding to its respective experimental aspect. Detailed methodologies for data acquisition and processing can be found in the original Science Advances article. ## Sharing/Access information The complete dataset is publicly accessible via Dryad: * [Dryad Repository](https://doi.org/10.5061/dryad.rn8pk0pn4) The data were generated as part of the experiments described in the article. For additional context and detailed experimental protocols, please refer to the original publication: * [Science Advances Article](https://www.science.org/doi/10.1126/sciadv.adj1896)