Caspase 1

The cortex tissue samples of the kidneys from each group were homogenized with protease inhibitors, lysis buffer, and a 1 mM mixture of phenylmethylsulfonyl fluoride. Total protein concentration was assayed using a bicinchoninic acid protein assay kit. Equal amounts of total protein extracts were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis at 20 μg per well and transferred to a polyvinylidene difluoride (PVDF) membrane. The PVDF membrane was immersed in tris-buffered saline Tween-20 with 3% bovine serum albumin for 1.5 h. The membrane was then incubated with the following specific primary antibodies (1:1000) overnight at 4°C: Anti-Keap1, anti-Nrf2, anti-Pro-caspase-1, anti-cleaved caspase-1, and anti-GSDMD. The membranes were then incubated with peroxidase-coupled anti-rabbit or anti-mouse antibodies for 2 h at room temperature. Super-enhanced chemiluminescence reagent (Applygen Technologies Inc, Beijing, China) was added to the membrane to visualize the target band. The band intensity was calculated using ImageJ software, and the relative protein intensities were calculated. Anti-Keap1, Nrf2, and caspase-1 were purchased from Servicebio (Wuhan, China), cleaved caspase-1 antibody was purchased from Santa Cruz (1:200, Dallas, TX, USA), and GSDMD and actin antibodies were obtained from Abcam (Cambridge, MA, USA).