Histological Analysis of Kidney Injury

The kidney tissues were perfused with 0.1 M phosphate-buffered saline (PBS) and fixed with 4% paraformaldehyde. They were further processed to embed them with paraffin and dissected into 7 µm thick slices. For the H&E staining, the sections were dehydrated with ethanol, stained with H&E, and mounted with Canada balsam (Kanto Chemical, Tokyo, Japan). For PAS staining, the PAS Stain Kit (Azer Scientific Inc., Morgantown, PA, USA) was used according to the manufacturer’s instructions. Histological glomerular injury was scored as follows according to the damaged regions by referring to previously established procedures [27 (link)]: 0: normal; 1: less than 25%; 2: between 25% and 50%; 3: between 50% and 75%; and 4: between 75% and 100%.