WB analyses was performed as described below. Initially, the radio-immunoprecipitation assay (RIPA) lysis buffer (Lot No:89,900; Beyotime, Jiangsu, China), which consisted of protease inhibitors and phosphatase inhibitors (Lot No: P1048; Beyotime, Jiangsu, China), was employed to lyse and extract the proteins present in the kidneys. Subsequently, the protein content was determined using the BCA method (Lot No: 23,227; Thermo Fisher, USA). Equivalent quantities of protein lysates were loaded and subjected to electrophoresis on a 10–12% sodium dodecyl sulfate polyacrylamide gel at a voltage of 100 V. Following this, the protein was transferred to a polyvinylidene fluoride membrane at a voltage of 120 V for a duration of 2 h. To prevent the binding of non-specific proteins, the membrane was treated with 5% non-fatty milk for 1 h at room temperature. Finally, the membrane was divided based on the protein’s kilodalton values. The membranes were subjected to overnight incubation at 4 °C with primary antibodies in order to obtain specific proteins. Subsequently, the membranes underwent a 1-hour incubation at room temperature with secondary antibodies labeled with horseradish peroxidase. Finally, the blot was detected and visualized using Clarity Western Enhanced chemiluminescence (ECL) reagent (Lot No: WBKLS0500; Millipore, USA) on an MP Imager (Bio-Rad, USA). Densitometric analysis of protein bands was conducted using Image-J (Bio-Rad, USA). The primary antibodies used in this experiment were as follows: Anti-NGAL antibody (1:1000, Lot No: ab41105) (Abcam, USA), Anti-SOD1 antibody (1:1000, Lot No: ab308181) (Abcam, USA), Anti-IL-10 antibody (1:1000, Lot No:ab34843) (Abcam, USA), Rabbit Anti-DRP1 (1:1000, Lot No: #8570s) (Cell Signaling Technology, USA), Rabbit Anti-Phospho-SAPK/JNK (1:1000, Lot No: #4668T) (Cell Signaling Technology, USA), Rabbit Anti-Phospho-RIP3 (1:1000, Lot No: #15828T) (Cell Signaling Technology, USA), Rabbit Anti-Phospho-MLKL (1:1000, Lot No: #37,333) (Cell Signaling Technology, USA), Anti-GAPDH (1:2000, Lot No: #2118) (Cell Signaling Technology, USA), Phospho-RIPK1 Monoclonal antibody (1:2000, Lot No:66854-1-Ig) (Proteintech, Wuhan, China), Phospho-ERK1/2 Polyclonal antibody (1:1000, Lot No:28733-1-AP) (Proteintech, Wuhan, China), ERK1/2 Polyclonal antibody (1:2000, Lot No: 11257-1-AP) (Proteintech, Wuhan, China), JNK Monoclonal antibody (1:3000, Lot No: 66210-1-Ig) (Proteintech, Wuhan, China), OPA1 Polyclonal antibody (1:1500, Lot No: 27733-1-AP) (Proteintech, Wuhan, China), MFF Polyclonal antibody (1:1000, Lot No: 17090-1-AP) (Proteintech, Wuhan, China), PGC-1α Monoclonal antibody (1:1000, Lot No:66369-1-Ig) (Proteintech, Wuhan, China), GAPDH Monoclonal antibody (1:5000, Lot No: 60004-1-Ig) (Proteintech, Wuhan, China), Anti-TNF-α (1:300, Lot No: sc-52,746) (Snata Cruz, USA), Anti-IL-6 (1:300, Lot No:sc-57,315) (Snata Cruz, USA), Anti-MCP-1 (1:300, Lot No: sc-52,701) (Snata Cruz, USA), and Mouse TIM-1/KIM-1/HAVCR Antibody (1:500, Lot No: AF1817) (R&D system, USA). MaxVision HRP-polymer anti-mouse/rabbit secondary antibodies (Lot No: KIT-5020; Maixin Biotech, Fujian, China) and Goat Anti-Rabbit IgG secondary antibody (Lot No: #L3012; Signalway, USA) were used in this experiment. The explanation of the original data of WB experiment was as follows. Three different samples per group were used for WB experiment to meet the requirements for biological replication [
28]. The membrane was cut according to the kilodaltons of specific proteins. The other parts of the same membrane were also kept. In this study, the original images of all-length cropped blots were presented in the form of supplementary file
3. And the regions of the original blots used in main figures were marked in red boxes.