Kidney tissues were paraffin-embedded sectioned, Each slide was dewaxed by xylene, ethanol treatment, and antigen repair with citric acid as the repair solution, after which it was closed with a serum of the same source as the secondary antibody (usually 10% goat serum was used) and incubated for 30 min at 37°C (Henkels et al., 2016 (link); Sasaki et al., 2021 (link)). Then treated slides with anti-Ly-6G (Servicebio; GB11229), anti-F4/80 antibodies (28463-1-AP, Proteintech, Wuhan, China) overnight at 4°C. All slides were washed with PBS before incubation with FITC-labeled goat anti-mouse antibody and CY3-labeled goat anti-mouse antibody (Pinuofei Biotechnology Co, China). After PBS rinsing, the slides were exposed to DAPI for 5 min to stain cell nuclei. Then, the slides were analyzed under a fluorescence microscope.
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