Cleaved caspase 1

H9C2 cells or rat myocardial tissues were harvested, and protein was extracted using RIPA lysis buffer (Thermo Fisher Scientific) that includes protease inhibitors (Roche Diagnostic, Indianapolis, IN, USA). The protein extraction process was kept on ice. The quantity of total protein was determined by BCA assay (Thermo Scientific). Protein samples (20 µg) were  loaded  and fractionated by 10% SDS-PAGE and then transferred to polyvinylidene difluoride membrane (Millipore, Billerica, MA, USA). Subsequently, membranes were incubated with 5% non-fat milk containing 0.1% PBST to block nonspecific binding for 2 h at room temperature and treated with primary antibodies against ELAVL1 (1:1000, Abcam, Cambridge, MA, USA), NLRP3 (1:1000, Abcam), cleaved caspase-1 (1:1000, Cell Signaling Technology, Danvers, MA, USA), and GAPDH (1:1000, Abcam) at 4℃ overnight, respectively, followed by incubation with the relevant horseradish peroxidase (HRP) conjugated secondary antibody (1:5000, Abcam) at room temperature for 2 h. The protein bands were visualized using an enhanced chemiluminescence kit (Synthgene) and quantified by analysis using the ImageJ software.
Exosome marker proteins CD9, CD63, and TSG101 were identified by western blot, the protocol being similar to that used for cell and tissue protein detection.

Dulbecco’s modified Eagle medium and penicillin–streptomycin solution were purchased from GIBCO (Grand Island, NY, USA). Fetal bovine serum was provided by Hyclone (Logan, UT, USA). Monoclonal rabbit antibodies, including anti-p38 MAPK, anti-phospho-p38 MAPK, anti-ERK1/2, anti-phospho-ERK1/2, anti-JNK, anti-phospho-JNK, anti-p65, anti-phospho-p65, NLRP3, and cleaved caspase-1 were obtained from Cell Signaling Technology (Boston, MA, USA). Receptor for AGE (RAGE) monoclonal rabbit antibody was purchased from AbCam (Cambridge, MA, USA). HRP-marked anti-β-actin antibody was supplied by Biorad (San Diego, CA, USA). 4′-Methoxyresveratrol (3,5-dyhydroxy-4′-methoxylstilbene) was purchased from Great Forest Biomedical (Hangzhou, China). BSA was obtained from ABCONE (Shanghai, China). KI and acetic acid were obtained from Sangon Biotech (Shanghai, China). Methylglyoxal, 2’,7’-dichlorodihydrofluorescein diacetate and other reagents were purchased from Sigma (St. Louis, MO, USA).