Protein Quantification and Western Blotting Protocol

Western blotting was conducted as previously described [8 (link)]. Similarly, we obtained the supernatant of the sample and extracted proteins using the radio immunoprecipitation assay (RIPA) buffer containing 1 mmol/L PMSF (Beyotime, Shanghai, China), and the protein concentration was quantified using a BCA assay kit (Beyotime, Shanghai, China). During the process of electrophoresis, the proteins with different sizes were separated in SDS-PAGE. Then, the proteins on the gel were transferred onto a polyvinylidene-difluoride (PVDF) membrane (Beyotime, Shanghai, China) for blotting. Antibodies were acquired from Beyotime Biotechnology, Shanghai, China, which included glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mouse monoclonal antibody (catalog number: AG019), caspase-1rabbit polyclonal antibody (catalog number: AF1681), Nrf2 rabbit polyclonal antibody (catalog number: AF7623), NF-κB p65 rabbit polyclonal antibody (catalog number: AF5875), HO-1rabbit polyclonal antibody (catalog number: AF1333), NLRP3 rabbit monoclonal antibody (catalog number: AF2155), HRP-labeled goat anti-rabbit IgG (H + L) (catalog number: A0208) and HRP-labeled goat anti-mouse IgG (H + L) (catalog number: A0216). Original Western Blot figures in Figure S2.