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.