Supplementary raw data of Berberine inhibits NLRP3 inflammasome activation by upregulating the SIRT6-AMPK pathway to prevent angiotensin II-induced fibrosis and vulnerability to atrial fibrillation
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Full unedited gel of this study. Blue boxes indicate the lanes corresponding to those shown in the cropped images within the manuscript
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Proteins were extracted from atrial tissue with RlPA lysis bufer, phenylmethyl sulfony fuoride (PMSF), and universaphosphatase 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 dodecy sulfate polyacrylamide gel electrophoresis (SDS-PAGF) was pertormed, 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 were diluted in PBST, The membrane was incubated with a primary antibody overnight on a shaking bed at 4℃, followed by washing with PBST and subsequent incubation with a secondary antibody at room temperature for 2 hours, Target bands were visualized using a Tanon-5200 imaging system (bioTanon, Shanghai, China).