Supplementary raw data of Reduced FNDC5-AMPK signaling in diabetic atrium increases the susceptibility of atrial fibrillation by impairing mitochondrial dynamics and activating NLRP3 inflammasome
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Full unedited gel of this study. Red boxes indicate the lanes corresponding to those shown in the cropped images within the manuscript.
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Proteins were extracted from mouse atrial tissue and cultured cells with RIPA lysis buffer, phenylmethyl sulfonyl fluoride (PMSF), and universal phosphatase inhibitor cocktail at a ratio of 100:1:1. The process was conducted on ice, followed by centrifugation at 12000 rpm for 20 minutes. Protein concentration was determined using the BCA kit (P0011, Beyotime Biotech, China), and the proteins were boiled for preservation. Subsequently, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was performed, and the proteins were transferred to a polyvinylidene fluoride (PVDF) membrane (7082493, GVS Group, USA). The membranes were blocked with 5% bovine serum albumin (BSA) in phosphate-buffered saline (PBS) containing 0.1% Tween-20 (PBST). Primary antibodies against were diluted in PBST. The membrane was incubated with a primary antibody overnight on a shaking bed at 4°C, followed by washing with PBST and subsequent incubation with a secondary antibody at room temperature for 2 hours. Protein bands were detected with an image analyzer.
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- Liaoning Revitalization Talents ProgramLiaoning, ChinaGrant ID: XLYC2203006
- Shenyang Science and Technology ProjectGrant ID: 22-321-33-36