Cryosections from frozen kidney tissues (5 μm) were prepared using a Leica CM1900 cryostat (Leica). As previously described [47 (link)], the sections were stained with dihydroethidium (DHE) solution for 30 min in the dark at 37 °C and then washed three times with phosphate-buffered saline (PBS). Finally, a fluorescence microscope (Olympus; Tokyo, Japan) was used to photograph the section.
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