Bs 4081r

Heart tissue sections were deparaffinized and rehydrated for 20 min with xylene and graded ethanol solution. Antigen retrieval was performed using boiling 0.01 M citrate buffer. Endogenous peroxidase activity was suppressed by incubation with 3% hydrogen peroxide. All sections were incubated with primary antibodies against KAT5 (bs-13686R, 1:100, Bioss, Beijing, China), STUB1 (ab134064, 1:100, Abcam, UK), LATS2 (bs-4081R, 1:100, Bioss), YAP (bs-3605R, 1:100, Bioss), β-catenin (A19657, 1:50, Abclonal), and NLRP3 (MA5-32255, 1:50, Thermo Fisher) overnight at 4 °C. After thorough washing, the sections were incubated with biotin-conjugated secondary antibody for 30 min and colored with di-amino benzidine. All sections were counterstained with hematoxylin and imaged using a light microscope.

RIPA buffer (Solarbio) was used for total protein extraction, and the protein concentration was assessed by BCA Protein Assay Kit (Cat. No.: PC0020, Solarbio). The protein samples were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, and transferred to polyvinylidene fluoride membranes, followed by blocking in 5% skim milk for 1 h. Subsequently, the membranes were probed with primary antibodies against KAT5 (bs-13686R, 1:500, Bioss), STUB1 (A11751, 1:1000, Abclonal, Wuhan, China), LATS2 (bs-4081R, 1:500, Bioss), p-YAP (bsm-52214R, 1:500, Bioss), YAP (bs-3605R, 1:1000, Bioss), β-catenin (A19657, 1:1000, Abclonal), NLRP3 (A5652, 1:500, Abclonal), ASC (bs-6741R, 1:500, Bioss), Caspase-1 (A0964, 1:1000, Abclonal), IL-18 (A1115, 1:500, Abclonal), IL-1β (A16288, 1:500, Abclonal), β-actin (bs-0061R, 1:5000, Bioss) overnight at 4 °C. After incubation with the secondary antibody for 1 h, the protein bands were developed by the ECL Western Blotting Substrate (Cat. No.: PE0010, Solarbio). For tissue detection, five samples from each group were used. For cell detection, three samples from three independent experiments were used.