A13014

Myocardial tissue was used to extract the samples, which were then boiled in a loading buffer for five minutes. 10% SDS-PAGE was used to separate the proteins. Primary antibodies were grown on the PVDF membranes overnight at 4 °C. The primary antibodies used in this study were Actin (Dilution 1:1000, M20011, Abmart), Pdk4 (Dilution 1:1000, YN5701, Immunoway), Hmgcs2 (Dilution 1:5000, ab137043, abcam), Decr1 (Dilution 1:1000, A13014, ABclonal), and Ivd (Dilution 1:2000, 10822-1-AP, ProteinTech). The membranes were incubated with the secondary antibody for 1 h at room temperature. Finally, the membranes were detected by the ECL system.

Dewaxed, rehydrated, and antigen retrieval were performed on paraffin slices. Methanol was used to inactivate endogenous peroxidase for 15 min. The slices were then sealed after being treated with 5% BSA for 1 h, followed by an overnight incubation with primary antibodies. The primary antibodies used in this study were Pdk4 (Dilution 1:50, YN5701, Immunoway), Hmgcs2 (Dilution 1:200, ab137043, abcam) Decr1 (Dilution 1:50, A13014, ABclonal) and Ivd (Dilution 1:500, 10822-1-AP, ProteinTech). The paraffin slices were cultured with secondary antibodies the next day for one hour at 37 °C. Finally, the slides were stained with a DAB Detection Kit, and the counterstained with haematoxylin.