Senescence-Associated β-Galactosidase Staining of Mouse Kidney

SA-β-Gal staining was performed using a previously published protocol (Debacq-Chainiaux et al., 2009 (link); Geng et al., 2019 (link); Zhang et al., 2021 (link)). In brief, OCT-embedded, snap-frozen, unfixed mouse kidney tissues were cryosectioned at a thickness of 15 μm with a Leica CM3050S cryomicrotome, collected on Superfrost Plus microslides (VWR) and kept at –80°C until use. For SA-β-Gal staining, sections were thawed at RT and processed by using Senescence β-Galactosidase Staining Kit (Beyotime, Cat#C0602) as the manufacturer’s recommendation. Images were taken with Carl Zeiss 200 microscope, and the SA-β-Gal-positive areas were quantified using ImageJ.