Cells and frozen renal tissue sections were fixed with 4% paraformaldehyde in PBS for 10 min at room temperature, washed with PBS, and permeabilized with 0.3% Triton X-100 for 10 min. After blocking in 1% BSA for 30 min, the cells were immunolabeled with primary antibodies (anti-Kim-1 at 1:150 dilution; anti-TNF-α at 1:100 dilution; anti-dsDNA at 1:400 dilution; anti-TOM20 at 1:100 dilution; or anti-TFAM at 1:50 dilution) overnight at 4 °C followed by incubation with FITC- or TRITC-conjugated secondary antibody (1:200) for 1 h at 37 °C. Nuclei were visualized by staining with DAPI for 5 min at room temperature. Digital images of the sections were captured using a fluorescence microscope (Nikon, N-STORM & A1, Tokyo, Japan), and the Pearson's correlation coefficient between the TFAM and dsDNA results was analyzed using Image-Pro Plus.

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