Interaction of KCNA5, CX43, and CX40 proteins in the atrial muscle of patients with atrial fibrillation

免疫沉淀 免疫荧光 免疫印迹 连接蛋白 污渍 信使核糖核酸 化学 分子生物学 细胞生物学 基因 生物化学 生物 缝隙连接 抗体 免疫学 细胞内
作者
Keke Wang,Juntao Zhao,Zhikun Guo
出处
期刊:Cell Biology International [Wiley]
卷期号:46 (11): 1834-1840 被引量:5
标识
DOI:10.1002/cbin.11864
摘要

The objective of the study was to investigate the expression levels of potassium voltage-gated channel subfamily A member 5 (KCNA5), connexin 43 (Cx43), and connexin 40 (Cx40) in the left atrial appendage of patients with atrial fibrillation (AF) and the interactions between them. We gathered tissue samples from patients with persistent AF and sinus rhythm and used fluorescence quantitative polymerase chain reaction to evaluate messenger RNA (mRNA) changes of KCNA5, Cx43, and Cx40. Then, we studied the protein levels of KCNA5, Cx43, and Cx40 by immunofluorescence and western blot analysis and the interactions between these proteins were identified by immunoprecipitation and immunofluorescence colocation, respectively. Compared with the control group, the mRNA and protein levels of KCNA5, Cx43, and Cx40 in the AF group were decreased and the positive expression of KCNA5, Cx43, and Cx40 protein was also decreased by immunofluorescence staining in the AF group. In addition, immunoprecipitation and immunofluorescence colocation revealed that KCNA5 was coexpressed with Cx43 and Cx40 proteins. The expressions of KCNA5, Cx43, and Cx40 were substantially downregulated in the myocardium of patients with AF and KCNA5 interacted with Cx43 and Cx40 proteins, respectively.
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