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目的:证实α-Actinin 2与小电导钙激活钾通道(Small Conductance Ca~(2+)-activated K~+Channels,SK)的共同表达增加SK2在HEK293细胞膜上的表达。方法:HEK293细胞用RPMI 1640培养基培养。将HEK293细胞分为两组,对照组细胞转染pIRES—SK2质粒,实验组细胞共转染pIRES—SK2和pcDNA3—α-actinin2质粒。免疫荧光共聚焦屈微镜和蛋白印迹技术检测SK2通道蛋白在HEK293细胞膜上的表达。结果:免疫荧光共聚焦显微镜检测到转染pIRES—SK2质粒的HEK293细胞膜上有SK2通道蛋白的表达。蛋白印迹技术显示实验组蛋白条带亮度明显高于对照组。结论:α—Actinin 2能增加SK2在HEK293细胞膜上的表达。
OBJECTIVE: To confirm the co-expression of α-actinin 2 and small Conductance Ca ~ (2 +) - activated K ~ + Channels, SK) on the membrane of HEK293 cells. Methods: HEK293 cells were cultured in RPMI 1640 medium. The HEK293 cells were divided into two groups. The control cells were transfected with pIRES-SK2 plasmid. The cells in the experimental group were co-transfected with pIRES-SK2 and pcDNA3-α-actinin2 plasmids. Immunofluorescence confocal microscopy and Western blotting were used to detect the expression of SK2 channel protein in HEK293 cell membrane. Results: Immunofluorescence confocal microscopy showed the expression of SK2 channel protein in HEK293 cell membrane transfected with pIRES-SK2 plasmid. Western blotting showed that the brightness of the experimental histone band was significantly higher than that of the control group. Conclusion: α-Actinin 2 can increase the expression of SK2 in HEK293 cell membrane.