Kidneys were fixed in 4% PFA for 24 hours and embedded in paraffin. Periodic acid–Schiff (PAS) staining was performed to evaluate pathological injury. Masson and Sirius red staining were performed to estimate renal fibrosis. Two blinded renal pathologists quantified staining in eight randomly selected fields. Data were analyzed using Image-Pro Plus software (Media Cybernetics, Rockville, MD, USA).

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