Anti nlrp3

Maresin‐1 and Maresin‐1 ELISA kit were purchased from Cayman Chemical. AngII was purchased from Enzo Life Sciences. Mouse IL‐1β ELISA kit was purchased from Neobioscience. EdU imaging kits was purchased from APExBIO. ML385 and KN‐93 were purchase from Topscience. Fluo‐4 AM was purchased from Invitrogen. Fetal bovine serum was purchased from Inner Mongolia Opcel Biotechnology Co., Ltd. Primary antibodies included: anti‐GAPDH (Abcolonal), anti‐SM22α (Servicebio), anti‐ASC (Santa Cruz Biotechnology), anti‐caspase‐1 p20 (Santa Cruz Biotechnology), anti‐IL‐1β (Santa Cruz Biotechnology), anti‐OPN (Immunoway), anti‐NLRP3 (Immunoway), anti‐IL‐18 (ImmunoWay), anti‐GSDMD‐N (Immunoway), anti‐Nrf2 (GeneTex), anti‐α‐SMA(Abcam), anti‐HO‐1(Abcam), anti‐CaMKII (Abcam), anti‐p‐CaMKII (Zenbio), anti‐PCNA (Cell Signaling Technology), anti‐rabbit IgG (H+L) (Cell Signaling Technology), anti‐mouse IgG (H+L) (Cell Signaling Technology), Cy3‐conjugated goat anti‐rabbit IgG (H+L) (Servicebio), and Alexa Fluor® 488‐conjugated goat anti‐rabbit IgG (H+L) (Servicebio).

First, cortical tissues were washed three times with PBS to remove the blood and cut into small pieces. Next, tissue samples were lysed and centrifuged, and the resulting supernatant was collected as the total protein solution. Total protein concentration was determined using a bicinchoninic acid (BCA) protein assay kit (Beyotime). The proteins were then separated using sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE), followed by transfer to polyvinylidene difluoride (PVDF) membranes. The membranes were blocked with 5% non‐fat dry milk in Tris‐buffered saline containing Tween 20 (TBST) and probed with primary antibodies overnight at 4°C, including anti‐NLRP3 (Immunoway), ASC (Immunoway), cleaved‐caspase‐1 (Immunoway), GSDMD‐N (Immunoway, United Kingdom), HMGB1 (Bioss), TLR4 (Bioss), NF‐κB p65 (Immunoway), p38 MAPK (Immunoway), JNK (Immunoway) and anti‐β‐actin (CST) antibodies. After washing three times with TBST, the membranes were incubated with the corresponding secondary antibodies (Servicebio), and the protein bands were detected using a Bio‐Rad Gel imaging system (Bio‐Rad). The quantification of protein bands was performed using Image J software, and the Western blot results were normalized to β‐actin expression.