Measurement of ROS levels in the kidney tissues

The levels of ROS were analyzed in the tissues utilizing the dihydroethidium (DHE) fluorescent probe (cat. no. D7008; MilliporeSigma), according to the manufacturer's protocol. The sections were stained by incubation with 50 µM DHE for 1 h in the dark at room temperature, and then stained with 1 mg/ml DAPI for 10 min at 4˚C. The sections were rinsed three times with PBS. A fluorescent microscope (magnification, x200) was used to evaluate the fluorescence intensity at an excitation wavelength of 525 nm and an emission wavelength of 610 nm.