2.8 Immunohistochemistry and enhanced Prussian blue staining
For immunohistochemistry analysis, the slides were sequentially incubated with 5% bovine serum albumin for 1 h, with a primary antibody targeting F4/80 overnight, and then with the appropriate secondary antibody. After incubation with the secondary antibody, the cells were observed under an optical microscope. Enhanced Prussian blue staining was used to detect the iron deposition in renal tissue. Mouse kidney tissues were fixed with 3% glutaraldehyde overnight, washed three times, and sectioned at a thickness of 4 μm for prussian blue staining. After washing with PBS, nuclei were stained with hematoxylin, and the iron deposition in the tissues was finally observed under an optical microscope.